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Repeated social defeat increases reactive emotional coping behavior and alters functional responses in serotonergic neurons in the rat dorsal raphe nucleus
Abstract
Chronic stress is a vulnerability factor for a number of psychiatric disorders, including anxiety and affective disorders. Social defeat in rats has proven to be a useful paradigm to investigate the neural mechanisms underlying physiologic and behavioral adaptation to acute and chronic stress. Previous studies suggest that serotonergic systems may contribute to the physiologic and behavioral adaptation to chronic stress, including social defeat in rodent models. In order to test the hypothesis that repeated social defeat alters the emotional behavior and the excitability of brainstem serotonergic systems implicated in control of emotional behavior, we exposed adult male rats either to home cage control conditions, acute social defeat, or social defeat followed 24h later by a second social defeat encounter. We then assessed behavioral responses during social defeat as well as the excitability of serotonergic neurons within the dorsal raphe nucleus using immunohistochemical staining of tryptophan hydroxylase, a marker of serotonergic neurons, and the protein product of the immediate-early gene, c-fos. Repeated social defeat resulted in a shift away from proactive emotional coping behaviors, such as rearing (explorative escape behavior), and toward reactive emotional coping behaviors such as freezing. Both acute and repeated defeat led to widespread increases in c-Fos expression in serotonergic neurons in the dorsal raphe nucleus. Changes in behavior following a second exposure to social defeat, relative to acute defeat, were associated with decreased c-Fos expression in serotonergic neurons within the dorsal and ventral parts of the mid-rostrocaudal dorsal raphe nucleus, regions that have been implicated in 1) serotonergic modulation of fear- and anxiety-related behavior and 2) defensive behavior in conspecific aggressive encounters, respectively. These data support the hypothesis that serotonergic systems play a role in physiologic and behavioral responses to both acute and repeated social defeat.
... We place special focus on the serotonin system and its 2A receptor (5-HT2AR) subtype in particular, which has been shown to be particularly implicated in biology x environment interactions (Chang et al., 2017;Dressler et al., 2016;Fiocco et al., 2007;Jiang et al., 2016;Jokela et al., 2007;Lebe et al., 2013;Mellman et al., 2009;Parade et al., 2017;Salo et al., 2011). Various acute stressors appear to be reliable and robust inducers of serotonin release (Adell et al., 1997;Amat et al., 1998Amat et al., , 2005Bastani et al., 2017;Beitia et al., 2005;Bland et al., 2003;Cohen et al., 2015;Ferres-Coy et al., 2013;Fuenmayor and Garcia, 1984;Fujino et al., 2002;Gardner et al., 2005;Hale et al., 2011;Higuchi et al., 2019;Ishida et al., 1997;Ishiwata and Greenwood, 2018;Johnson et al., 2005;Keeney et al., 2006;Kelly et al., 2011;Li et al., 2015;Myers and Beleslin, 1971;Nakajima et al., 2009;Neugebauer, 2020;Paul et al., 2011;Rex et al., 2005;Yoshioka et al., 1995) and different types of stressor, such as sleep deprivation (Elmenhorst et al., 2012;Maple et al., 2015;Zhao et al., 2019), hypoxia (Anju et al., 2011), chronic tryptophan depletion (Cahir et al., 2007), inflammation (Zhang et al., 2001), tonic pain (Kupers et al., 2009), repeated forced swim (Takao et al., 1995), repeated shock (Dwivedi et al., 2005), administration of stress hormones (Jitsuiki et al., 2000;Kuroda et al., 1992Kuroda et al., , 1994, amygdala stimulation (Kalynchuk et al., 2006), time-dependent stress , novelty stress (Aloyo and Dave, 2007), maternal separation (Benekareddy et al., 2010(Benekareddy et al., , 2011Godar et al., 2019;Vazquez et al., 2000; although see Ohta et al., 2014), isolation rearing (Preece et al., 2004;Rilke et al., 1998), isolation housing (Günther et al, 2008;Schiller et al., 2003;although see Bibancos et al., 2007;Schiller (Berton et al., 1997;McKittrick et al., 1995; although see Visser et al., 2014) all appear to upregulate 5-HT2AR expression, particularly in the cerebral cortex (Anju et al., 2011;Benekareddy et al., 2010Benekareddy et al., , 2011Berton et al., 1997;Cahir et al., 2007;Dwivedi et al., 2005;Elmenhorst et al., 2012;Godar et al., 2019;Günther et al, 2008;Harvey et al., 2003;Jitsuiki et al., 2000;Kalynchuk et al., 2006;Kupers et al., 2009;Kuroda et al., 1992Kuroda et al., , 1994Maple et al., 2015;McKittrick et al., 1995;Ossowska et al., 2001;Preece et al., 2004;Rilke et al., 1998;Takao et al., 1995;Vazquez et al., 2000;Zhao et al., 2019; see also Table 1), which is well known to be massively expanded in humans. Social isolation and defeat also reliably sensitize behavioural responses to 5-HT2AR agonists (Benekareddy et al., 2010;Brotto et al., 1998;Gorzalka et al., 1998;Sakaue et al., 2002;Sood et al., 2018;Wright et al., 1991; see also Table 1). ...
... In preclinical modelling studies, 5-HT2AR antagonism impairs acquisition of conditioned defeat (Clinard et al., 2015;Rillich and Stevenson, 2018) and suppresses hyperthermic response to social defeat (Beig et al., 2009;Sinh and Ootsuka, 2019), whereas agonist administration into the basolateral amygdala increases acquisition of conditioned defeat (Clinard et al., 2015). Acute social defeat increases 5-HT levels (Beitia et al., 2005;Gardner et al., 2005;Higuchi et al., 2019;Keeney et al., 2006;Paul et al., 2011), and region-specific increases in 5-HT2AR expression have been found in chronically subordinate animals (Berton et al., 1997;McKittrick et al., 1995). Acute social defeat has not been shown to upregulate 5-HT2AR expression (Visser et al., 2014) or produce phenotypes indicative of 5-HT2AR sensitization, such as that seen after recurring defeat (Hayashida et al., 2010). ...
... As discussed in previous sections, certain types of stress upregulate 5-HT2AR expression and activity (see Table 1 for a review of the relevant literature). Moreover, evidence that acute stress causes the release of 5-HT is compelling (Adell et al., 1997;Amat et al., 1998Amat et al., , 2005Bastani et al., 2017;Beitia et al., 2005;Bland et al., 2003;Cohen et al., 2015;Ferres-Coy et al., 2013;Fuenmayor and Garcia, 1984;Fujino et al., 2002;Gardner et al., 2005;Hale et al., 2011;Higuchi et al., 2019;Ishida et al., 1997;Ishiwata and Greenwood, 2018;Johnson et al., 2005;Keeney et al., 2006;Kelly et al., 2011;Li et al., 2015;Myers and Beleslin, 1971;Nakajima et al., 2009;Neugebauer, 2020;Paul et al., 2011;Rex et al., 2005;Yoshioka et al., 1995) and it is logical to surmise that increased endogenous 5-HT release will engage the 5-HT2AR system, an assumption backed up by evidence of increased 5-HT2AR associated responses with drug-induced 5-HT release (Kuypers et al., 2018;Liechti et. al., 2000;Pitts et al., 2017;van Wel et al., 2012). ...
This paper introduces a new construct, the ‘pivotal mental state’, which is defined as a hyper-plastic state aiding rapid and deep learning that can mediate psychological transformation. We believe this new construct bears relevance to a broad range of psychological and psychiatric phenomena. We argue that pivotal mental states serve an important evolutionary function, that is, to aid psychological transformation when actual or perceived environmental pressures demand this. We cite evidence that chronic stress and neurotic traits are primers for a pivotal mental state, whereas acute stress can be a trigger. Inspired by research with serotonin 2A receptor agonist psychedelics, we highlight how activity at this particular receptor can robustly and reliably induce pivotal mental states, but we argue that the capacity for pivotal mental states is an inherent property of the human brain itself. Moreover, we hypothesize that serotonergic psychedelics hijack a system that has evolved to mediate rapid and deep learning when its need is sensed. We cite a breadth of evidences linking stress via a variety of inducers, with an upregulated serotonin 2A receptor system (e.g. upregulated availability of and/or binding to the receptor) and acute stress with 5-HT release, which we argue can activate this primed system to induce a pivotal mental state. The pivotal mental state model is multi-level, linking a specific molecular gateway (increased serotonin 2A receptor signaling) with the inception of a hyper-plastic brain and mind state, enhanced rate of associative learning and the potential mediation of a psychological transformation.
... The DR neurons are highly responsive to different environmental stimuli, including stressful experiences or antidepressant treatments (Grandjean et al., 2019;Kawahara et al., 1993;Kelly et al., 2011;Nishii et al., 2017). When persistent, environmental challenges can induce adaptive modifications in the DR neurophysiology resulting in altered serotonergic transmission (Lo Iacono et al., 2019;Paul et al., 2011; https://doi.org/10.1016/j.brainres.2020.146763 Received 4 December 2019; Received in revised form 3 March 2020; Accepted 8 March 2020 T Popa et al., 2010;Wood et al., 2013). ...
... Received 4 December 2019; Received in revised form 3 March 2020; Accepted 8 March 2020 T Popa et al., 2010;Wood et al., 2013). These neuroplastic responses can have a high impact on behavioral outputs and are implicated in a variety of psychopathological conditions (Lo Iacono et al., 2019;Paul et al., 2011;Prakash et al., 2019;Wood et al., 2013). Indeed, chronic stress-induced serotonergic dysregulation is known to be highly involved in the pathogenesis of disorders, including anxiety, mood disorders and addiction (Christianson and Greenwood, 2014;Hale et al., 2012;Lupien et al., 2009;Muller and Homberg, 2015). ...
... The DR is highly responsive to environmental challenges like stress exposure. Following repeated stimulation, the DR can undergo neuroplastic modifications that can alter its excitability and the kinetics of 5-HT release in target structures persistently (Adell et al., 1988;Bambico et al., 2009;Paul et al., 2011;Wood et al., 2013). In this study we applied a repeated restraint stress manipulation to induce an adaptive response in the mouse DR (Chung et al., 2000). ...
The Dorsal Raphe (DR) is the primary source of serotonergic input in the brain and a center for the homeostatic maintenance of the serotonergic tone. Under repeated stimulation, it can undergo adaptive modifications that alter serotonergic neurotransmission, which can lead to behavioral dysfunction.
Post-transcriptional regulation by microRNAs is implicated in these adaptations. However, a global microRNA/target network effect on the DR neuroplasticity has yet to be elucidated. Here we investigate the microRNAs/mRNAs regulatory activity in the mouse DR after a chronic stress experience.
First, we assessed the behavioral consequences of repeated restraint stress exposure and the functional adaptations of the DR by measuring the change in acute stress-induced serotonin release. Then, through next generation RNA-Seq of Argonaute2-bound RNA (RISC-Seq) we identified microRNAs and their targets that are associated to the RISC complex of the DR in unstressed and stressed mice.
We mapped the potential microRNA/mRNA network within the stress-altered transcripts, uncovering new interactions that contribute to the chronic stress-induced DR modifications.
... After a single exposure to SD, rats had an increased Fos expression in CRH 2 positive neurons in the mAMY. 56 The role of the mAMY in stress response may be associated with defeat behaviors, such as avoidance of aggressive levels of c-Fos in the PFC and DR brain areas 63,71 and changes in histones and calcium/calmodulin-dependent kinase in the hippocampus. 48,69 In a more comprehensive description, Carboni et al. 64 reported changes in foldingrelated proteins, signal transduction, synaptic plasticity, cytoskeleton regulation and energy metabolism in the hippocampus after repeated psychosocial stress exposure. ...
... 59,60 The serotonergic system might be involved in inhibitory processes of proactive coping responses, such as aggression 121,122 and escape behaviors 123 ; at the same time, it may be associated with facilitation of passivesubmissive responses. 124 Using a repeated SD stress protocol, Paul et al. 63 found a shift away from a proactive emotional coping style and towards a reactive emotional coping style during the defeat phase. In unstable social structure periods, adopting a subordinate position may prevent dangerous situations and limit injury and energy wastage. ...
... days had more 22 kHz USV, 70 more passive coping behaviors,23,63,70 increased anxiety-like behaviors, such as increased self-grooming, increased locomotion in novel environments and risk assessment behavior,23,51,64,65,72 and increased depressive-like symptoms, such as reduced sweet solution preference and intake, reduced climbing in the FST, lower general activity and sociability in the social avoidance test and reduced mounting in copulatory behavior.48,51,66,71 Repeatedly stressed LR rats displayed social avoidance under infusion of BDNF antagonist in hippocampus, whereas repeatedly stressed HR rats displayed social approach under infusion of BDNF agonist in hippocampus.46 ...
Social defeat (SD) in rats, which results from male intraspecific confrontations, is ethologically relevant and useful to understand stress effects on physiology and behavior.
A systematic review of studies about biomarkers induced by the SD protocol and published from 2002 to 2013 was carried out in the electronic databases PubMed, Web of Knowledge and ScienceDirect. The search terms were: social defeat, rat, neurotrophins, neuroinflammatory markers, and transcriptional factors.
Classical and recently discovered biomarkers were found to be relevant in stress-induced states. Findings were summarized in accordance to the length of exposure to stress: single, repeated, intermittent and continuous SD. This review found that the brain-derived neurotrophic factor (BDNF) is a distinct marker of stress adaptation. Along with glucocorticoids and catecholamines, BDNF seems to be important in understanding stress physiology.
The SD model provides a relevant tool to study stress response features, development of addictive behaviors, clinic depression and anxiety, as well as individual differences in vulnerability and resilience to stress.
... During acutely distressful experiences, greater vmPFC engagement is seen in stress-resilient humans (Sinha et al., 2016) and non-human animals (Baratta et al., 2009;Morrison et al., 2014), whereas reduced activation corresponds with vulnerability to stress-induced impairments. Similarly, several acutely administered traumatic stressors stimulate serotonergic activity within DRN, though in an opposing manner with activation correlating positively with stress susceptibility (Amat et al., 2005;Paul et al., 2011;Cooper et al., 2017). Taken together, these findings indicate that an inverse relationship between vmPFC and DRN activity coincides with resistance to the adverse effects of acute traumatic stress. ...
... GABAergic interneurons within the lateral wings of the DRN, which then synapse onto and inhibit serotonin-producing cells along the DRN's midline (e.g., Jankowski and Sesack, 2004;Challis et al., 2014). Given that acute stress-induced activation of serotonergic cells is positively correlated with stress susceptibility (Amat et al., 2005;Cooper et al., 2008Cooper et al., , 2017Paul et al., 2011), activation of vmPFC-DRN projections is thought to functionally suppress the DRN's response during acute stress and buffer its negative consequences. Indeed, vmPFC-DRN cell recruitment is associated with indices of resilience to non-social laboratory stressors such as tail shock (Baratta et al., 2009(Baratta et al., , 2019, elevated plus maze (Faye et al., 2020), and forced swim (Warden et al., 2012). ...
The ventromedial prefrontal cortex (vmPFC) plays a critical role in stress resilience through top-down inhibition of key stress-sensitive limbic and hindbrain structures, including the dorsal raphe nucleus (DRN). In a model of experience-dependent stress resistance, socially dominant Syrian hamsters display fewer signs of anxiety following acute social defeat when compared to subordinate or control counterparts. Further, dominants activate vmPFC neurons to a greater degree during stress than do subordinates and become stress-vulnerable following pharmacological inhibition of the vmPFC. Dominants also display fewer stress-activated DRN neurons than subordinates do, suggesting that dominance experience gates activation of vmPFC neurons that inhibit the DRN during social defeat stress. To test whether social dominance alters stress-induced activity of a vmPFC-DRN pathway, we injected a retrograde tracer, cholera toxin B (CTB), into the DRN of dominant, subordinate, and control hamsters and used a dual-label immunohistochemical approach to identify vmPFC neurons co-labeled with CTB and the defeat-induced expression of an immediate early gene, cFos. Results indicate that dominant hamsters display more cFos+ and dual-labeled cells in layers V/VI of infralimbic and prelimbic subregions of the vmPFC compared to other animals. Furthermore, vmPFC-DRN activation corresponded directly with proactive behavioral strategies during defeat, which is indicative of stress resilience. Together, results suggest that recruiting the vmPFC-DRN pathway during acute stress corresponds with resistance to the effects of social defeat in dominant hamsters. Overall, these findings indicate that a monosynaptic vmPFC-DRN pathway can be engaged in an experience-dependent manner, which has implications for behavioral interventions aimed at alleviating stress-related psychopathologies.
... We lastly examined behaviors of oxytocin receptordeficient mice during social defeat stress to clarify physiological roles of the oxytocin receptor in the control of defensive behaviors during social defeat stress. Social defeat stress was applied twice because it has been shown that experience of social defeat induces social memory and facilitates defensive behavior in the second social interaction (49)(50)(51)(52) and that the oxytocin receptor plays a role in the acquisition of social memory (33). ...
... On the other hand, the duration of freezing behavior during the second stress was increased compared with that during the first stress, consistent with the results of a previous study (49), and there was no significant difference between wild-type mice and oxytocin receptordeficient mice [repeated-measures two-way ANOVA (F 1,31 = 1.780; P = 0.203; no significant effects of genotype), (F 1,31 = 5.736; P = 0.0312; significant effects of number of times for stress), (F 1,31 = 0.0739; P = 0.790; no significant interaction)] [ Fig. 7(b)]. ...
Social stress has deteriorating effects on various psychiatric diseases. In animal models, exposure to socially dominant conspecifics, i.e., social defeat stress, evokes species-specific defeat posture via unknown mechanisms. Oxytocin neurons have been shown to be activated by stressful stimuli and to have pro-social and anxiolytic actions. The roles of oxytocin during social defeat stress remain unclear. Expression of c-Fos, a marker of neuronal activation, in oxytocin neurons and in oxytocin receptor-expressing neurons was investigated in mice. The projection of oxytocin neurons was examined with an anterograde viral tracer, which induces selective expression of membrane-targeted palmitoylated green fluorescent protein in oxytocin neurons. Defensive behaviors during double exposure to social defeat stress in oxytocin receptor-deficient mice were analyzed. Following social defeat stress, expression of c-Fos protein was increased in oxytocin neurons of the bed nucleus of the stria terminalis, supraoptic nucleus, and paraventricular hypothalamic nucleus. Expression of c-Fos protein was also increased in oxytocin receptor-expressing neurons of brain regions including the ventrolateral part of the ventromedial hypothalamus and ventrolateral periaqueductal gray. Projecting fibers from paraventricular hypothalamic oxytocin neurons were found in the ventrolateral part of the ventromedial hypothalamus and in the ventrolateral periaqueductal gray. Oxytocin receptor-deficient mice showed reduced defeat posture during the second social defeat stress. These findings suggest that social defeat stress activates oxytocin-oxytocin receptor systems, and the findings are consistent with the view that activation of the oxytocin receptor in brain regions including the ventrolateral part of the ventromedial hypothalamus and the ventrolateral periaqueductal gray facilitates social defeat posture.
... CRF-induced motor activity (Zorrilla et al. 2002). Low motor activity of adolescent rats injected with CP376395 could explain the reduced rates of attacks and the increased latency to attack toward these less active intruders, since reduced intruder locomotion is thought to reduce resident aggressive behavior (Buwalda et al. 2012;Paul et al. 2011). As the residents spent less time attacking the CP376395-infused intruders they spent more time sniffing them, suggesting residents were no less interested in the drug-treated intruders, just less aggressive toward them. ...
... For example, lower levels of motor activity during the social defeat test are thought to reduce further aggression when the intruder is an adolescent Fig. 4 Top, latency to adopt the supine posture during the fourth defeat on P44 was negatively correlated with the number of cocaine infusions obtained during the 24-h binge for rats infused with aCSF prior to social defeat stress and those infused with CP376395. Bottom, latency to supine on P44 also was inversely correlated with cocaine infusions obtained during progressive ratio schedule of reinforcement (van den Berg et al. 1999) and when it is an adult (Buwalda et al. 2012;Nocjar et al. 2012;Paul et al. 2011). It is not possible to determine whether the increase in aggression from P35 to P44 is a result of the conditioned behavior of the intruder or the increased probability of attacking a larger and older adolescent intruder undergoing sexual maturation (Burke and Miczek 2014). ...
Background
Activation of corticotropin-releasing factor type 1 receptors (CRF-R1) in the ventral tegmental area (VTA) represents a critical mechanism for social defeat to escalate cocaine self-administration in adult rats.
Objective
We determined the acute effect of a CRF-R1 antagonist (CP376395) microinfusion into the VTA prior to each episode of social defeat in adolescent rats and determined whether this drug treatment could prevent later escalation of cocaine taking in early adulthood.
Methods
Rats were implanted with bilateral cannulae aimed at the VTA 5 days before the first social defeat. Bilateral microinfusion of CP376395 (500 ng/side) or vehicle occurred 20 min before each episode of social defeat on postnatal days (P) 35, 38, 41, and 44. Behavior was quantified on P35 and P44. On P57, rats were implanted with intra-jugular catheters, and subsequent cocaine self-administration was analyzed.
Results
CP376395-treated adolescent rats walked less and were attacked more slowly but were socially investigated more than vehicle-treated adolescents. Vehicle-treated rats showed increased social and decreased non-social exploration from P35 to P44, while CP376395-treated rats did not. Socially defeated, vehicle-treated adolescents took more cocaine during a 24-h unlimited access binge during adulthood. The latency to supine posture on P44 was inversely correlated with later cocaine self-administration during fixed and progressive ratio schedules of reinforcement and during the binge.
Conclusions
CP376395 treatment in adolescence blocked escalation of cocaine taking in adulthood. Episodes of social defeat stress engender neuroadaptation in CRF-R1s in the VTA that alter coping with social stress and that persist into adulthood.
... Losing also failed to modify the DRN 5-HT response to IS, with elevated 5-HT levels persisting throughout the entire shock session. It should be noted that other acute stressors, such as social defeat, activate DRN 5-HT cells (Cooper et al., 2009;Amat et al., 2010;Paul et al., 2011) and produce similar behavioral outcomes as does IS such as social avoidance and shuttlebox escape failure . This defeat-induced DRN 5-HT activation is selectively suppressed in animals with dominant status (Cooper et al., 2017). ...
Dominance status has extensive effects on physical and mental health, and an individual’s relative position can be shaped by experiential factors. A variety of considerations suggest that the experience of behavioral control over stressors should produce winning in dominance tests and that winning should blunt the impact of later stressors, as does prior control. To investigate the interplay between competitive success and stressor control, we first examined the impact of stressor controllability on subsequent performance in a warm spot competition test modified for rats. Prior experience of controllable, but not physically identical uncontrollable, stress increased later effortful behavior and occupation of the warm spot. Controllable stress subjects consistently ranked higher than did uncontrollable stress subjects. Pharmacological inactivation of the prelimbic (PL) cortex during behavioral control prevented later facilitation of dominance. Next, we explored whether repeated winning experiences produced later resistance against the typical sequelae of uncontrollable stress. To establish dominance status, triads of rats were given five sessions of warm spot competition. The development of stable dominance was prevented by reversible inactivation of the PL or NMDA receptor blockade in the dorsomedial striatum. Stable winning blunted the later stress-induced increase in dorsal raphe nucleus serotonergic activity, as well as prevented uncontrollable stress-induced social avoidance. In contrast, endocrine and neuroimmune responses to uncontrollable stress were unaffected, indicating a selective impact of prior dominance. Together, these data demonstrate that instrumental control over stress promotes later dominance, but also reveal that winning experiences buffer against the neural and behavioral outcomes of future adversity.
... Stress also influences 5-HT 107 neurotransmission (Graeff et al., 1996;Hale et al., 2012). Alterations in brain 5-HT, 5-HT-related 108 molecular content (Kang et al., 2005), and serotonergic activity (Paul et al., 2011;Zhang et al., 2012), 109 have been observed in individuals exposed to acute or chronic stress. In behavioral experiments, 110 mice with hereditary central 5-HT deficiency are more susceptible to social stress (Sachs et al., 2015), 111 and increasing 5-HT levels with SSRIs prevents the impairment of stress adaptation (Uno et al.,112 2019). ...
Depression is a frequent and serious illness, and stress is considered the main risk factor for its onset. First-line antidepressants increase serotonin (5-hydroxytryptamine; 5-HT) levels in the brain. We previously reported that an N -acetyltransferase, Shati/Nat8l, is upregulated in the dorsal striatum (dSTR) of stress-susceptible mice exposed to repeated social defeat stress (RSDS) and that dSTR Shati/Nat8l overexpression in mice (dSTR-Shati OE) induces stress vulnerability and local reduction in 5-HT content. Male mice were used in this study, and we found that dSTR 5-HT content decreased in stress-susceptible but not in resilient mice. Moreover, vulnerability to stress in dSTR-Shati OE mice was suppressed by the activation of serotonergic neurons projecting from the dorsal raphe nucleus (dRN) to the dSTR, followed by upregulation of 5-HT content in the dSTR using designer receptors exclusively activated by designer drugs (DREADD). We evaluated the role of GABA in modulating the serotonergic system in the dRN. Stress-susceptible after RSDS and dSTR-Shati OE mice exhibited an increase in dRN GABA content. Furthermore, dRN GABA content was correlated with stress sensitivity. We found that the blockade of GABA signaling in the dRN suppressed stress susceptibility in dSTR-Shati OE mice. In conclusion, we propose that dSTR 5-HT and dRN GABA, controlled by striatal Shati/Nat8l via the dSTR-dRN neuronal circuitry, critically regulate stress sensitivity. Our study provides insights into the neural processes that underlie stress and suggests that dSTR Shati/Nat8l could be a novel therapeutic target for drugs against depression, allowing direct control of the dRN serotonergic system.
... ; https://doi.org/10.1101/2023.06.06.543982 doi: bioRxiv preprint elevated 5-HT levels persisting throughout the entire shock session. It should be noted that other acute stressors, such as social defeat, activate DRN 5-HT cells (Cooper et al., 2009;Amat et al., 2010;Paul et al., 2011) and produce similar behavioral outcomes as does IS such as social avoidance and shuttlebox escape failure (Amat et al., 2010). ...
Dominance status has extensive effects on physical and mental health, and an individual's relative position can be shaped by experiential factors. A variety of considerations suggest that the experience of behavioral control over stressors should produce winning in dominance tests and that winning should blunt the impact of later stressors, as does prior control. To investigate the interplay between competitive success and stressor control, we first examined the impact of stressor controllability on subsequent performance in a warm spot competition test modified for rats. Prior experience of controllable, but not physically identical uncontrollable, stress increased later effortful behavior and occupation of the warm spot. Controllable stress subjects consistently ranked higher than did uncontrollable stress subjects. Pharmacological inactivation of the prelimbic (PL) cortex during behavioral control prevented later facilitation of dominance. Next, we explored whether repeated winning experiences produced later resistance against the typical sequelae of uncontrollable stress. To establish dominance status, triads of rats were given five sessions of warm spot competition. Reversible inactivation of the PL or NMDA receptor blockade in the dorsomedial striatum led to a long-term reduction in social rank. Stable dominance blunted the later stress-induced increase in dorsal raphe nucleus serotonergic activity, as well as prevented stress-induced social avoidance. In contrast, endocrine and neuroimmune responses to uncontrollable stress were unaffected, indicating a selective impact of prior dominance. Together, these data demonstrate that instrumental control over stress promotes later dominance, but also reveal that winning experiences buffer against the neural and behavioral outcomes of future adversity.
... In fact, optogenetic stimulation of DRN neurons and 5-HTergic projections from the DRN to the nucleus accumbens can even rescue autism-like social deficits in mice (Luo et al., 2015;Walsh et al., 2018). Additionally, social behavior modifications following uncontrollable stress and aversive social situations like social defeat are mediated by 5-HT action in the DRN, basolateral amygdala, and dorsal striatum (Christianson et al., 2010;Cooper et al., 2008;Lukkes et al., 2009;Paul et al., 2011;Strong et al., 2011). ...
Social interaction allows for the transfer of affective states among individuals, and the behaviors and expressions associated with pain and fear can evoke anxiety-like states in observers which shape subsequent social interactions. We hypothesized that social reactions to stressed individuals engage the serotonergic dorsal raphe nucleus (DRN) which promotes anxiety-like behavior via postsynaptic action of serotonin at serotonin 2C (5-HT2C) receptors in the forebrain. First, we inhibited the DRN by administering an agonist (8-OH-DPAT, 1ug in 0.5uL) for the inhibitory 5-HT1A autoreceptors which silences 5-HT neuronal activity via G-protein coupled inward rectifying potassium channels. 8-OH-DPAT prevented the approach and avoidance, respectively, of stressed juvenile (PN30) or stressed adult (PN50) conspecifics in the social affective preference (SAP) test in rats. Similarly, systemic administration of a 5-HT2C receptor antagonist (SB242084, 1mg/kg, i.p.) prevented approach and avoidance of stressed juvenile or adult conspecifics, respectively. Seeking a locus of 5-HT2C action, we considered the posterior insular cortex which is critical for social affective behaviors and rich with 5-HT2C receptors. SB242084 administered directly into the insular cortex (5uM bilaterally in 0.5uL) interfered with the typical approach and avoidance behaviors observed in the SAP test. Finally, using fluorescent in situ hybridization, we found that 5-HT2C receptor mRNA (htr2c) is primarily colocalized with mRNA associated with excitatory glutamatergic neurons (vglut1) in the posterior insula. Importantly, the results of these treatments were the same in male and female rats. These data suggest that interactions with stressed others require the serotonergic DRN and that serotonin modulates social affective decision-making via action at insular 5-HT2C receptors.
... One possibility is that the pharmacological blockade of mGlu7 may affect the dorsal raphe nucleus serotonin system and connected forebrain circuits, which are key central systems that are supposed to modulate anxiety-and stress-related behaviors and determine behavioral coping responses. This may result in a shift from reactive to proactive coping during CSC exposure [41][42][43]. Importantly, there is clear evidence from human and rodent studies that the way of behavioral coping predicts stress vulnerability and that there is a lowered risk to develop stress-related affective, as well as somatic diseases in proactively coping individuals [44,45]. However, with respect to pharmacological mGlu7 blockade and CSC exposure, this possibility is so far just speculative and has to be addressed experimentally in future studies. ...
Chronic psychosocial stress participates prominently in the etiology of various psychiatric conditions and comorbid somatic pathologies; however, suitable pharmacotherapy of these disorders is still of high medical need. During the last few decades, research on mGlu receptors advanced remarkably and much attention was given to the mGlu7 subtype. Here, genetic mGlu7 ablation, short-term pharmacological mGlu7 blockade, as well as siRNA-mediated knockdown of mGlu7 were shown to result in an acute anti-stress, antidepressant- and anxiolytic-like phenotype in mice. Moreover, we recently revealed a prominent stress-protective effect of genetic mGlu7 ablation also with respect to chronic psychosocial stress. In addition, we are able to demonstrate in the present study that the chronic pharmacological blockade of mGlu7 interferes with various chronic stress-induced alterations. For this, we used the chronic subordinate colony housing (CSC), a mouse model of chronic male subordination, in combination with chronic treatment with the mGlu7-selective orthosteric-like antagonist XAP044 (7-hydroxy-3-(4-iodophenoxy)-4H-chromen-4-one). Interestingly, XAP044 dose-dependently ameliorates hypothalamic–pituitary–adrenal axis dysfunctions, thymus atrophy, as well as the CSC-induced increase in innate anxiety. Taken together, our findings provide further evidence for the role of mGlu7 in chronic psychosocial stress-induced alterations and suggests the pharmacological blockade of mGlu7 as a promising therapeutic approach for the treatment of chronic stress-related pathologies in men.
... One possibility is that the pharmacological blockade of mGlu7 may affect the dorsal raphe nucleus serotonin system and connected forebrain circuits, which are key central systems that are supposed to modulate anxiety-and stress-related behaviors and determine behavioral coping responses. This may result in a shift from reactive to proactive coping during CSC exposure [41][42][43]. Importantly, there is clear evidence from human and rodent studies that the way of behavioral coping predicts stress vulnerability and that there is a lowered risk to develop stress-related affective, as well as somatic diseases in proactively coping individuals [44,45]. However, with respect to pharmacological mGlu7 blockade and CSC exposure, this possibility is so far just speculative and has to be addressed experimentally in future studies. ...
Chronic psychosocial stress participates prominently in the etiology of various psychiatric conditions and comorbid somatic pathologies; however, suitable pharmacotherapy of these disorders is still of high medical need. During the last few decades, research on mGlu receptors advanced remarkably and much attention was given to the mGlu7 subtype. Here, genetic mGlu7 ablation, short-term pharmacological mGlu7 blockade, as well as siRNA-mediated knockdown of mGlu7 were shown to result in an acute anti-stress, antidepressant- and anxiolytic-like phenotype in mice. Moreover, we recently revealed a prominent stress-protective effect of genetic mGlu7 ablation also with respect to chronic psychosocial stress. In addition, we are able to demonstrate in the present study that the chronic pharmacological blockade of mGlu7 interferes with various chronic stress-induced alterations. For this, we used the chronic subordinate colony housing (CSC), a mouse model of chronic male subordination, in combination with chronic treatment with the mGlu7-selective orthosteric-like antagonist XAP044 (7-hydroxy-3-(4-iodophenoxy)-4H-chromen-4-one). Interestingly, XAP044 dose-dependently ameliorates hypothalamic–pituitary–adrenal axis dysfunctions, thymus atrophy, as well as the CSC-induced increase in innate anxiety. Taken together, our findings provide further evidence for the role of mGlu7 in chronic psychosocial stress-induced alterations and suggests the pharmacological blockade of mGlu7 as a promising therapeutic approach for the treatment of chronic stress-related pathologies in men.
... The 5-HT system is thus ideally placed to support integration of emotional, motivational and cognitive behaviors [22,23]. For example, in animal models, 5-HT in limbic forebrain areas was altered by multiple environmental stressful conditions, such as temperature variation [24], swim stress [25], or social defeat [26]. In humans, patients with anxiety and affective disorders show an increase of brain 5-HT turnover compared to healthy subjects [27,28]. ...
... Thus, these two studies [3,47] suggest that LiCl-induced toxicosis, but not LPS-induced sickness, increases conditioned rearing responses. It has been suggested that increases in vertical activity are a measure of attempted escape behavior from an aversive environment, rather than an increase in exploratory behavior [10,40,[46][47][48]. Unsupported rearing (in the center of an open-field) also seems to be a stable measure of anxiety induced escape behavior [46]. ...
Lithium chloride (LiCl) is an emetic drug that has been used to create animal models of anticipatory nausea and conditioned place aversion. In this study we examined escape behaviours from a context in which rats experienced the aversive effects of LiCl treatments. The experiment had two phases: acquisition of context conditioning, which consisted of pairing a distinct context with the pharmacological effects of a moderate dose of the toxin LiCl, and extinction of context conditioning, which consisted of placement in the distinct context in a drug free state. During context conditioning, 16 adult male Long-Evans rats were injected intraperitoneally with 96 mg/kg lithium chloride (LiCl; n = 8) or 0.9% saline (NaCl; n = 8) and placed individually in an automated locomotor activity apparatus for 30 minutes every other day for 4 days. During the extinction phase, rats were placed in the apparatus for 30 minutes every other day without injections during a 4 day extinction phase. A significant Drug x Trial interaction was found for the time spent in vertical position in the open field apparatus during trials 1 - 3 of the extinction phase. The LiCl treated rats exhibited significantly increased rearing behavior, relative to the control rats, indicative of conditioned aversion. The results of this study suggest that escape behaviour (vertical activity) occurs in rats experiencing the aversive conditioned effects of LiCl in a distinct context. In the context of current theoretical accounts, the LiCl-conditioned increase in apparent escape behaviors can be considered a reflection of anticipatory nausea.
... The 5-HT system is thus ideally placed to support integration of emotional, motivational and cognitive behaviors [22,23]. For example, in animal models, 5-HT in limbic forebrain areas was altered by multiple environmental stressful conditions, such as temperature variation [24], swim stress [25], or social defeat [26]. In humans, patients with anxiety and affective disorders show an increase of brain 5-HT turnover compared to healthy subjects [27,28]. ...
We explored here the hypothesis that temporary chronic water restriction in mice affects social behavior, via its action on the density of 5-HT neurons in dorsal and median raphe nuclei (DRN and MRN). For that, we submitted adult C57BL/6 J mice to mild and controlled temporary dehydration, i.e., 6 h of water access every 48 h for 15 days. We investigated their social behavior in a social interaction task known to allow free and reciprocal social contact. Results showed that temporary dehydration increases significantly time spent in social contact and social dominance. It also expands 5-HT neuron density within both DRN and MRN and the behavioral and neuronal plasticity were positively correlated. Our findings suggest that disturbance in 5-HT neurotransmission caused by temporary dehydration stress unbalances choice processes of animals in social context.
... Les tests d'agressivité chez le rongeur classiquement utilisés en laboratoire incluent le modèle du résidentintrus (resident-intruder, RI), dans lequel un intrus non familier, sélectionné pour sa faible agressivité, est inséré dans la cage de la souris expérimentale, ainsi que le paradigme gagnant-perdant, dans lequel deux souris précédemment isolées établissent des contacts sensoriels au travers d'un séparateur transparent et perforé, qui est retiré régulièrement afin de permettre des contacts physiques directs (Paul et al., 2011 ;Krishnan et al., 2007 ;Mooney et al., 2014). Dans de tels tests d'établissement hiérarchique, la corrélation entre l'agressivité et le rang social dans le tube n'est pas toujours observée. ...
L'organisation hiérarchique est la marque de nombreux vertébrés. Des associations entre rang social et état de santé sont signalées chez de nombreuses espèces incluant l’humain. Cependant, les mécanismes impliqués sont mal compris. La réponse au stress, spécifique au rang social chez de nombreux animaux, intervient dans de nombreuses pathologies psychiatriques. Ceci suggère un rôle de la réponse au stress dans le processus liant statut social et comportement. Mon travail de thèse vise à mieux comprendre les associations qui existent entre le rang social et le comportement individuel chez la souris. Mon travail vise à clarifier la direction de cette association en observant les conséquences du rang social sur le comportement et la vulnérabilité aux maladies psychiatriques, et réciproquement, en identifiant des marqueurs individuels précoces pouvant contribuer à façonner le destin social. Au niveau physiologique, je me concentre sur la réponse au stress et ses conséquences sur le système dopaminergique (DA), en tant que médiateur potentiel des phénotypes sociaux. Pour répondre à ces questions, j'ai évalué le rang social de mâles adultes C57BL/6 via des tests compétitifs. J'ai ensuite testé ces animaux pour l'anxiété, la sociabilité et les compétences cognitives, avant et après établissement hiérarchique. Nous avons enregistré l'activité électrophysiologique de cellules DA dans la région tégmentale ventrale. J'ai quantifié la libération de DA dans la voie mésocorticolimbique et testé les souris pour la dépression et la dépendance. Enfin, j’ai évalué les conséquences d’une inactivation du récepteur des glucocorticoïdes dans les cellules DAceptives pour la dominance sociale.
... Previously it was shown that rats that resist to defeat by showing defensive behaviors are less vulnerable to the lasting consequences of social defeat stress (28). It seems that WTGs exhibit a lot more 'proactive behaviors', while Wistars exhibit a lot more 'reactive behaviors' (29,30). Even though the WTG rats seem to cope better with the social defeat experience, the speed of recovery of the neuroendocrine stress response is delayed in comparison with that in Wistar rats. ...
Suppression of hippocampal neurogenesis is a readout for stress-induced alterations in neuroplasticity. In this study, we hypothesized that a single episode of severe social or non-social stress would differentially suppress neurogenesis in the dentate gyrus (DG) 10 days later in two rat strains. We anticipated that the suppression following social stress would be less severe in wildtype Groningen (WTG) rats, a rat strain considered relatively resilient to social stressors. Male Wistar and WTG were subjected to either social defeat or to immobilization stress. Behavioral response to social defeat and acute corticosterone response to both stressors was measured as well as anxiety behavior 10 days later on the elevated plus maze. Subsequently, brains were collected following cardiac aldehyde perfusion. The behavioral freezing response to defeat was much stronger in Wistar rats as compared to WTG rats. Acute corticosteroid response was similar in both strains although Wistar rats more rapidly resumed baseline values. There was no significant effect of both stressors on hippocampal DG cell proliferation and differentiation as well as on anxiety behavior. However, a striking strain difference appeared in anxiety behavior and both markers of neurogenesis. The WTG strain exhibiting much lower anxiety as well as reduced rate of hippocampal neurogenesis under all treatments. The results in this study suggest that both short-lasting acute stressors failed to induce lasting anxiety or decreased neurogenesis in the DG. Future studies could explore if and how rate of hippocampal neurogenesis is related with behavioral coping with stress.
... Knock-out mice lacking key components of serotonergic machinery show altered stress-related behaviors (Holmes et al., 2003;Lira et al., 2003;Adamec et al., 2006;Gutknecht et al., 2015). Extracellular levels of serotonin (Kawahara et al., 1993;Mokler et al., 2007), expression of serotonin-related molecules (Adell et al., 1988;Zhang et al., 2012;Issler et al., 2014;Donner et al., 2018), serotonergic activity (Grahn et al., 1999;Paul et al., 2011;Grandjean et al., 2019), and innervation (Natarajan et al., 2017) have been shown to change in response to both acute and chronic stress. Such regulation is linked to behavioral changes including anhedonia (Berton et al., 1997;Wood et al., 2013;Lopes et al., 2016;Natarajan et al., 2017) and is altered by treatment with antidepressants and anxiolytics (Benmansour et al., 1999;Abumaria et al., 2007). ...
Chronic stress induces anhedonia in susceptible but not resilient individuals, a phenomenon observed in humans as well as animal models, but the molecular mechanisms underlying susceptibility and resilience are not well understood. We hypothesized that the serotonergic system, which is implicated in stress, reward and antidepressant therapy, may play a role. We found that plasticity of the serotonergic system contributes to the differential vulnerability to stress displayed by susceptible and resilient animals. Stress-induced anhedonia was assessed in adult male rats using social defeat and intracranial self-stimulation (ICSS), while changes in serotonergic phenotype were investigated using immunohistochemistry and in situ hybridization. Susceptible, but not resilient, rats displayed an increased number of neurons expressing the biosynthetic enzyme for serotonin, tryptophan-hydroxylase-2 (TPH2), in the ventral subnucleus of the dorsal raphe nucleus (DRv). Further, a decrease in the number of DRv glutamatergic (VGLUT3+) neurons was observed in all stressed rats. This neurotransmitter plasticity is activity-dependent, as was revealed by chemogenetic manipulation of the central amygdala, a stress-sensitive nucleus that forms a major input to the DR. Activation of amygdalar corticotropin releasing hormone (CRH)+ neurons abolished the increase in DRv TPH2+ neurons and ameliorated stress-induced anhedonia in susceptible rats. These findings show that activation of amygdalar CRH+ neurons induces resilience, and suppresses the gain of serotonergic phenotype in the DR that is characteristic of susceptible rats. This molecular signature of vulnerability to stress-induced anhedonia and the active nature of resilience could be targeted to develop new treatments for stress-related disorders like depression.SIGNIFICANCE STATEMENTDepression and other mental disorders can be induced by chronic or traumatic stressors. However, some individuals are resilient and do not develop depression in response to chronic stress. A complete picture of the molecular differences between susceptible and resilient individuals is necessary to understand how plasticity of limbic circuits is associated with the pathophysiology of stress-related disorders. Using a rodent model, our study identifies a novel molecular marker of susceptibility to stress-induced anhedonia, a core symptom of depression, and a means to modulate it. These findings will guide further investigation into cellular and circuit mechanisms of resilience, and the development of new treatments for depression.
... This finding might indicate a change or transition in the ANA coping strategy towards a more reactive strategy. Moreover, Paul et al. (2011) reported that repeated social defeat stress results in a change from proactive coping behaviors to reactive coping behaviors. Our results also showed changes in behavior after repeated exposure to defeat stress in all groups (a decrease in non-social exploration and an increase in immobility), which reduced the individual differences observed in the first confrontation. ...
The aims of this study were to identify behavioral strategies to cope with social defeat, evaluate their impact on tumor development and analyze the contributions of both to changes in physiology and behavior produced by chronic defeat stress. For this purpose, OF1 mice were inoculated with B16F10 melanoma cells and subjected to 18 days of repeated defeat stress in the presence of a resident selected for consistent levels of aggression. Combined cluster and discriminant analyses of behavior that manifested during the first social interaction identified three types of behavioral profiles: active/aggressive (AA), passive/reactive (PR) and an intermediate active/non-aggressive (ANA) profile. Animals that showed a PR coping strategy developed more pulmonary metastases at the end of the social stress period than animals in other groups. The ANA but not AA group also showed higher tumor metastases than non-stressed subjects. In addition, the ANA group differed from the other groups because it displayed the highest corticosterone levels after the first interaction. Chronic stress reduced sucrose consumption, which indicates anhedonia, in all the stressed groups. However, the PR subjects exhibited a longer immobility time and swam for less time than other subjects in the forced swim test (FST), and they travelled a shorter distance in the open field test (OFT). In this test, the ANA group also travelled smaller distances than the non-stressed group, but the difference was more moderate. In contrast, tumor development but not stress increased behaviors associated with anxiety in the OFT (e.g., time in the center) in all tumor-bearing subjects. In summary, although the effects of social stress and tumor development on behavior were rather moderate, the results indicate the importance of behavioral coping strategies in modulating the effects of chronic stress on health.
... Other studies have examined these and other nonsocial behaviors in relation to substance use and found that initial exposure to nicotine increased ambulation, but decreased rearing in adolescent rats (Schochet, Kelley, and Landry, 2004). The cocaine administered to the demonstrator may have increased rearing and self-grooming behaviors, which would, in turn, change the dynamics of the social interaction session (Paul et al., 2011). Thus, rearing and self-grooming in the observer was likely a shift to nonsocial exploratory and possibly escape behavior through social modeling in the observer. ...
Background
Drinking alcohol is facilitated by social interactions with peers, especially during adolescence. The importance of peer social influences during adolescence on alcohol and substance use has recently received more attention. We have shown that social interaction with an alcohol‐intoxicated peer influences adolescent alcohol drinking differently in male and female rats using the demonstrator–observer paradigm. The present set of experiments analyzed the social interaction session to determine changes in social behaviors and subsequent alcohol drinking in adolescent male and female rats.
Methods
Specifically, in Experiment 1, we determined whether specific social behaviors were altered during interaction with an alcohol‐intoxicated demonstrator administered 1.5 g/kg ethanol (EtOH) and assessed changes in EtOH intake in adolescent observers. Experiment 2 examined changes in voluntary saccharin consumption to determine whether social interaction with an alcohol‐intoxicated demonstrator administered 1.5 g/kg EtOH altered consumption of a palatable solution. In Experiment 3, we administered saline, and a low (5 mg/kg) or high (20 mg/kg) dose of cocaine to the demonstrator and assessed changes in the adolescent observers to determine whether social interaction with a “drugged” peer altered social behaviors and voluntary EtOH intake.
Results
We showed that social interaction with an alcohol‐intoxicated demonstrator administered 1.5 g/kg EtOH (i) decreased social play and increased social investigation and social contact in adolescent male and female observers, (ii) did not alter nonsocial behaviors, (iii) did not alter saccharin consumption, and (iv) increased voluntary EtOH intake in adolescent female but not male observers. When the peer was injected with cocaine, (i) social play was dose‐dependently decreased, (ii) there were no changes in other social or nonsocial behaviors, and (iii) voluntary EtOH intake in adolescent male and female observers was unaffected.
Conclusions
The present results are consistent and extend our previous work, showing that social interaction with an alcohol‐intoxicated peer selectively alters social behaviors and alcohol drinking in adolescent rats. Females appear to be more sensitive to the elevating effects of social interaction on voluntary EtOH consumption.
... This is based on previous work (Varlinskaya, Spear and Spear, 2001), where they indicated that the first 10 min of the trial was sufficient to characterize the entire social interaction trial. It is important to note that previous studies have also identified rearing as a proactive emotional coping behavior, specifically explorative escape behavior (Paul et al., 2011). ...
Background
Drinking alcohol is facilitated by social interactions with peers, especially during adolescence. The importance of peer social influences during adolescence on alcohol and substance use have recently received more attention. We have shown that social interaction with an alcohol-intoxicated peer influences adolescent alcohol drinking differently in male and female rats using the demonstrator-observer paradigm. The present set of experiments analyzed the social interaction session to determine behaviors that influence alcohol drinking in adolescent male and female rats.
Methods
Specifically, in experiment one we determined which behaviors were altered during social interaction with an alcohol-intoxicated demonstrator and assessed changes in ethanol intake in adolescent observers. Experiment two examined changes in voluntary saccharin consumption to determine if social interaction with an alcohol-intoxicated demonstrator altered consumption of a palatable solution. In experiment three, we administered a low (5 mg/kg) or high (20 mg/kg) dose of cocaine to the demonstrator and assessed changes in the adolescent observers to determine if social interaction with a ‘drugged’ peer altered social behaviors and voluntary ethanol intake.
Results
We showed that social interaction with an alcohol-intoxicated demonstrator (1) decreased social play and increased social investigation and social contact in adolescent male and female observers, (2) did not alter non-social behaviors, (3) did not alter saccharin consumption and (4) increased voluntary ethanol intake in adolescent female but not male observers. When the peer was injected with cocaine (1) social play was dose-dependently decreased, (2) there were no changes in other social or non-social behaviors, and (3) voluntary ethanol intake in adolescent male and female observers was unaffected.
Conclusions
The present results are consistent and extend our previous work showing that social interaction with an alcohol-intoxicated peer selectively alters social behaviors and alcohol-drinking in adolescent rats. Females appear to be more sensitive to elevating effects of social interaction on voluntary ethanol consumption.
... Knockout mice lacking key components of serotonergic machinery show altered 80 stress-related behaviors ( Holmes et al., 2003;Lira et al., 2003;Adamec et al., 2006;Gutknecht 81 et al., 2015). Extracellular levels of serotonin ( Kawahara et al., 1993;Mokler et al., 2007), 82 expression of serotonin-related molecules (Adell et al., 1988;Zhang et al., 2012;Issler et al., 83 2014; Donner et al., 2018), serotonergic activity ( Grahn et al., 1999;Paul et al., 2011;Grandjean 84 et al., 2019) and innervation ( Natarajan et al., 2017) have been shown to change in response to 85 both acute and chronic stress. Such regulation is linked to behavioral changes including 86 anhedonia ( Berton et al., 1997;Wood et al., 2013;Lopes et al., 2016;Natarajan et al., 2017) and 87 is altered by treatment with antidepressants and anxiolytics (Benmansour et al., 1999; 88 Abumaria et al., 2007). ...
Chronic stress induces anhedonia in susceptible, but not resilient individuals, a phenomenon observed in humans as well as animal models, but the molecular mechanisms underlying susceptibility and resilience are not well understood. We hypothesized that the serotonergic system, which is implicated in stress, reward and antidepressant therapy, may play a role. We found that plasticity of the serotonergic system contributes to the differential vulnerability to stress displayed by susceptible and resilient animals. Stress-induced anhedonia was assessed in adult male rats using social defeat and intracranial self-stimulation (ICSS), while changes in serotonergic phenotype were investigated using immunohistochemistry and in situ hybridization. Susceptible, but not resilient, rats displayed an increased number of neurons expressing the biosynthetic enzyme for serotonin, tryptophan-hydroxylase-2 (TPH2), in the ventral subnucleus of the dorsal raphe nucleus (DRv). Further, a decrease in the number of DRv glutamatergic neurons was observed in all stressed animals. This neurotransmitter plasticity is dependent on DR activity, as was revealed by chemogenetic manipulation of the central amygdala, a stress-sensitive nucleus that forms a major input to the DR. Activation of amygdalar corticotropin releasing hormone (CRH)+ neurons abolished the increase in DRv TPH2+ neurons and ameliorated stress-induced anhedonia in susceptible animals. These findings show that activation of amygdalar projections induces resilience, and suppresses the gain of serotonergic phenotype in the DR that is characteristic of susceptible animals. This molecular signature of vulnerability to stress-induced anhedonia and the active nature of resilience could be a target of new treatments for stress-related disorders like depression.
SIGNIFICANCE STATEMENT
Depression and other mental disorders can be induced by chronic or traumatic stressors. However, some individuals are resilient and do not develop depression in response to chronic stress. A complete picture of the molecular differences between susceptible and resilient individuals is necessary to understand how plasticity of limbic circuits is associated with the pathophysiology of stress-related disorders. Using a rodent model, our study identifies a novel molecular marker of susceptibility to stress-induced anhedonia, a core symptom of depression, and a means to modulate it. These findings will guide further investigation into cellular and circuit mechanisms of resilience, and the development of new treatments for depression.
... Inhibited rats who display withdrawal for novel social and nonsocial stimuli [24,27] are considered a useful animal model to study anxiety vulnerability [23,28], and research shows that these inhibited rats show an enhanced reactivity to stress [29]. In addition, behavioral inhibition in rats occurs as a response to repeated social defeat [30], and may cause early death due to greater physiological stress responses [16]. These exaggerated stress responses, as indicated by increased cortisol release in the hypothalamic-pituitary axis found in inhibited rats [31], is similar to findings from children with an inhibited temperament [32]. ...
... A number of anxiety-and stress-related stimuli have been shown to activate the dorsomedial DR (but not other subsets of serotonergic neurons, such as those in the DRVL/ VLPAG). This includes anxiogenic drugs such as the partial inverse agonist at the benzodiazepine allosteric site on the GABA A receptor, N-methyl-9H-pyrido [5,4-b]indole-3-carboxamide (FG-7142) (Abrams et al., 2005), the non-selective 5-hydroxytryptamine 2C (5-HT 2C ) receptor agonist m-chlorophenyl piperazine (m-CPP) (Abrams et al., 2005), anxiety-related neuropeptides (Hale, Stamper, et al., 2010;Staub et al., 2006Staub et al., , 2005, acoustic startle (Spannuth et al., 2011), and stress-or anxiety-related stimuli (Bouwknecht et al., 2007;Cordero et al., 2012;Gardner et al., 2005;Paul et al., 2011;Spiacci et al., 2012). The selective activation of the dorsomedial DR may be due to activation of afferents that selectively impact dorsomedial DR serotonergic neurons. ...
Caffeine is the most commonly used drug in the world. However, animal studies suggest that chronic consumption of caffeine during adolescence can result in enhanced anxiety-like behavioral responses during adulthood. One mechanism through which chronic caffeine administration may influence subsequent anxiety-like responses is through actions on brainstem serotonergic systems. In order to explore potential effects of chronic caffeine consumption on brainstem serotonergic systems, we evaluated the effects of a 28-day exposure to chronic caffeine (0.3 g/L; postnatal day 28–56) or vehicle administration in the drinking water, followed by 24 h caffeine withdrawal, and subsequent challenge with caffeine (30 mg/kg; s.c.) or vehicle in adolescent male rats. In Experiment 1, acute caffeine challenge induced a widespread activation of serotonergic neurons throughout the dorsal raphe nucleus (DR); this effect was attenuated in rats that had been exposed to chronic caffeine consumption. In Experiment 2, acute caffeine administration profoundly decreased tph2 and slc22a3 mRNA expression throughout the DR, with no effects on htr1a or slc6a4 mRNA expression. Chronic caffeine exposure for four weeks during adolescence was sufficient to decrease tph2 mRNA expression in the DR measured 28 h after caffeine withdrawal. Chronic caffeine administration during adolescence did not impact the ability of acute caffeine to decrease tph2 or slc22a3 mRNA expression. Together, these data suggest that both chronic caffeine administration during adolescence and acute caffeine challenge during adulthood are important determinants of serotonergic function and serotonergic gene expression, effects that may contribute to chronic effects of caffeine on anxiety-like responses.
... Inhibited behavioral responses to threat are a major component of temperament in nonhuman primates (Chun and Capitanio, 2016;Rogers et al., 2008) and rodents (Castro et al., 2012). In rats, for example, repeated social defeat causes an increase in behavioral inhibition (Paul et al., 2011) which, in turn, has been related to greater physiological stress reactivity and early death (Cavigelli and McClintock, 2003). Behavioral inhibition in children is associated with difficulties in peer interactions, more social anxiety and loneliness, and increased susceptibility to adverse physiological and behavioral effects of perceived social stress (Fox et al., 2005;Kagan et al., 1988;Rapee, 2002;Rubin et al., 2009). ...
Background: Social inhibition may promote emotional problems in children, but little is known about this disposition in adults. Our research builds on a theory-based model to suggest that adult social inhibition involves distinct behavioral (inhibition), cognitive (sensitivity), and affective (withdrawal) characteristics. Methods: A total of 1385 adults completed measures of social inhibition, emotional distress, and social stress. Factor analyses, reliability estimates and regression analyses were used to examine the robustness of our model, and the validity of the 15-item Social Inhibition Questionnaire (SIQ15). Results: In Study 1 (N = 1180; Mage 46.9 years; 52% women), factor analysis confirmed that behavioral inhibition, interpersonal sensitivity, and social withdrawal reflected distinct facets of social inhibition. Next, we developed the SIQ15 that covers these facets with 5 items each; e.g. has difficulty making contact; expects negative reactions from others; keeps others at a distance. Study 2 (N = 209; Mage 20.3 years; 77% women) showed that the SIQ15 and its 5-item Inhibition, Sensitivity and Withdrawal facet scales were internally consistent (Cronbach's α between 0.86/0.94) and stable over time (test–retest between r = 0.73/0.78). The SIQ15 facets differentially predicted related inhibition (Behavioral Inhibition Scale), rumination (Penn State Worry Questionnaire) and withdrawal (Personality Inventory for DSM-5) scores at 6 months follow-up. Younger age and having no partner were associated with more social inhibition. Limitations: Findings are based on self-report; experimental and prospective studies are needed to further validate our inhibition model. Conclusions: Inhibition, sensitivity, and withdrawal are distinct manifestations of adult social inhibition that can be reliably assessed with the SIQ15. Research needs to examine how this multidimensional nature of social inhibition has an effect on stress, health, and wellbeing.
... Moreover, reports have delineated differential effects of fluoxetine, ketamine, and imipramine on gene expression in the mPFC, NAc, amygdala, and vHPC Vialou et al., 2015). Social defeat in rats resulted in a significant increase in cfos reactivity in serotonergic neurons in the DRN, suggesting that they play a critical role mediated social defeat-induced depressive-like behaviors (Paul et al., 2011). Tryptophan hydroxylase 2 knockin mice which display a 60-80% reduction in brain serotonin levels were more susceptible to CSDS and resistant to antidepressant treatment (Sachs et al., 2015). ...
Major depressive disorder (MDD) is a common but serious neuropsychiatric affliction that comprises a diverse set of symptoms such as the inability to feel pleasure, lack of motivation, changes in appetite, and cognitive difficulties. Given the patient to patient symptomatic variability in MDD and differing severities of individual symptoms, it is likely that maladaptive changes in distinct brain areas may mediate discrete symptoms in MDD. The advent and recent surge of studies using viral-genetic approaches have allowed for circuit-specific dissection of networks underlying motivational behavior. In particular, areas such as the ventral tegmental area (VTA), nucleus accumbens (NAc), and ventral pallidum (VP) are thought to generally promote reward, with the medial prefrontal cortex (mPFC) providing top-down control of reward seeking. On the contrary, the lateral habenula (LHb) is considered to be the aversive center of the brain as it has been shown to encode negative valence. The behavioral symptoms of MDD may arise from a disruption in the reward circuitry, hyperactivity of aversive centers, or a combination of the two. Thus, gaining access to specific circuits within the brain and how separate motivational-relevant regions transmit and encode information between each other in the context of separate depression-related symptoms can provide critical knowledge towards symptom-specific treatment of MDD. Here, we review published literature emphasizing circuit- and cell type-specific dissection of depressive-like behaviors in animal models of depression with a particular focus on the chronic social defeat stress model of MDD.
... Here, we confirmed this difference by showing that social status controls exhibit greater c-Fos immunoreactivity following restraint compared to social defeat. Likewise, acute social defeat in both rats and hamsters produces robust c-Fos expression in the DRN with modest habituation after repeated social defeat [59,60]. We found that all subjects, regardless of social status, showed increased c-Fos immunoreactivity in the cdDRN following restraint compared to social defeat. ...
Understanding the cellular mechanisms that control resistance and vulnerability to stress is an important step toward identifying novel targets for the prevention and treatment of stress-related mental illness. In Syrian hamsters, dominant and subordinate animals exhibit different behavioral and physiological responses to social defeat stress, with dominants showing stress resistance and subordinates showing stress vulnerability. We previously found that dominant and subordinate hamsters show different levels of defeat-induced neural activity in brain regions that modulate coping with stress, although the extent to which status-dependent differences in stress vulnerability generalize to non-social stressors is unknown. In this study, dominant, subordinate, and control male Syrian hamsters were exposed to acute physical restraint for 30 min and restraint-induced c-Fos immunoreactivity was quantified in select brain regions. Subordinate animals showed less restraint-induced c-Fos immunoreactivity in the infralimbic (IL), prelimbic (PL), and ventral medial amygdala (vMeA) compared to dominants, which is consistent with the status-dependent effects of social defeat stress. Subordinate animals did not show increased c-Fos immunoreactivity in the rostroventral dorsal raphe nucleus (rvDRN), which is in contrast to the effects of social defeat stress. These findings indicate that status-dependent changes in neural activity generalize from one stressor to another in a brain region-dependent manner. These findings further suggest that while some neural circuits may support a generalized form of stress resistance, others may provide resistance to specific stressors.
... The increased freezing exhibited by pair-housed rats suggests an adaptive coping style that agrees with a previous report that adolescent pair-housed rats exposed to adolescent social defeat are quicker to show submission from the first to the last defeat (Watt et al., 2009). For adult rats, the reduction in locomotion over repeated social defeats is attributed to increased freezing in the presence of the resident and is thought to reduce the probability of further attacks (Buwalda et al., 2012;Nocjar et al., 2012;Paul et al., 2011), which suggests that the pair-housed adolescent's increase in freezing from the first to fourth defeat is more similar to adult observations than is the reduction in freezing observed in the single housed adolescents. ...
Negative social experiences during adolescence are central features for several stress-related mental illnesses. Social play fighting behavior in rats peaks during early adolescence and is essential for the final maturation of brain and behavior. Manipulation of the rat adolescent social experience alters many neurobehavioral measurements implicated in anxiety, depression, and substance abuse. In this review, we will highlight the importance of social play and the use of three separate social stress models (isolation-rearing, social defeat, and social instability stress) to disrupt the acquisition of this adaptive behavior. Social stress during adolescence leads to the development of anxiety and depressive behavior as well as escalated drug use in adulthood. Furthermore, sex- and age-dependent effects on the hormonal stress response following adolescent social stress are also observed. Finally, manipulation of the social experience during adolescence alters stress-related neural circuits and monoaminergic systems. Overall, positive social experiences among age-matched conspecifics during rat adolescence are critical for healthy neurobehavioral maturation.
... Neural activation patterns in two bi-directionally connected regions known for modulating the behavioral response to stress, the dorsal raphe and the amygdala, were measured in HR/LR rats following the defensive burying task. Expression of c-Fos, and other immediate early genes, have previously been used to identify brain-regions and cell types that contribute to stress coping behavior [25][26][27][28]. For example, c-Fos mapping identified the A2 noradrenergic cell group as hyporesponsive to stress in Wistar-Kyoto rats, which display high levels or reactive coping and immobility on the forcedswim test. ...
Chronic stress triggers a variety of physical and mental health problems, and how individuals cope with stress influences risk for emotional disorders. To investigate molecular mechanisms underlying distinct stress coping styles, we utilized rats that were selectively-bred for differences in emotionality and stress reactivity. We show that high novelty responding (HR) rats readily bury a shock probe in the defensive burying test, a measure of proactive stress coping behavior, while low novelty responding (LR) rats exhibit enhanced immobility, a measure of reactive coping. Shock exposure in the defensive burying test elicited greater activation of HR rats’ caudal dorsal raphe serotonergic cells compared to LRs, but lead to more pronounced activation throughout LRs’ amygdala (lateral, basolateral, central, and basomedial nuclei) compared to HRs. RNA-sequencing revealed 271 mRNA transcripts and 33 microRNA species that were differentially expressed in HR/LR raphe and amygdala. We mapped potential microRNA-mRNA networks by correlating and clustering mRNA and microRNA expression and identified networks that differed in either the HR/LR dorsal raphe or amygdala. A dorsal raphe network linked three microRNAs which were down-regulated in LRs (miR-206-3p, miR-3559-5p, and miR-378a-3p) to repression of genes related to microglia and immune response (Cd74, Cyth4, Nckap1l, and Rac2), the genes themselves were up-regulated in LR dorsal raphe. In the amygdala, another network linked miR-124-5p, miR-146a-5p, miR-3068-3p, miR-380-5p, miR-539-3p, and miR-7a-1-3p with repression of chromatin remodeling-related genes (Cenpk, Cenpq, Itgb3 bp, and Mis18a). Overall this work highlights potential drivers of gene-networks and downstream molecular pathways within the raphe and amygdala that contribute to individual differences in stress coping styles and stress vulnerabilities.
... The dorsal raphe nucleus (DRN), as the origin of serotonergic projections to forebrain regions including the amygdala (Amy), helps regulate anxiety states and anxiety-related behavior (Campbell and Merchant, 2003;Graeff et al., 1996;Maier et al., 1993). Stimuli related to anxiety activate DRN 5-HT neurons (Commons, 2008;Hale et al., 2012;Kelly et al., 2011;Paul et al., 2011;Spannuth et al., 2011), so evaluating DRN serotonergic activation is important when studying the effects of toxicant substances on anxiety. ...
Diesel exhaust consists of diesel exhaust particles (DEPs) and gaseous compounds. Because previous research suggested that in utero exposure to DEPs affected spatial learning and memory in male offspring, while epidemiological evidence suggested disturbances in affect after prenatal exposure to particulates, we hypothesized that DEP exposure during pregnancy might also disturb affect. Here, we explored the effects of in utero exposure to DEPs on anxiety in male ICR mice. DEP solutions were administered subcutaneously to pregnant ICR mice at a dose of 0 or 200 μg/kg body weight on gestation days 6, 9, 12, 15, and 18. We assessed anxiety in 6 week-old male offspring using the hole board test and elevated plus maze test. After the behavioral tests, animals were sacrificed and serotonin (5-HT) levels in the dorsal raphe nucleus (DRN) were measured using HPLC. Mice exposed to DEPs in utero demonstrated increased anxiety in both behavioral tests. HPLC analysis revealed a significant increase in 5-HT levels in the DRN. Double immunolabeling of the DRN using anti-5-HT and anti-FosB (a chronic neuronal activation marker) antibodies indicated chronic activation of the DRN might underlie the increased anxiety after prenatal DEP exposure.
... These observations suggest that neurochemical adaptations occur in 5-HT neurons in response to environmental or pharmacological stressors. This is supported by studies in rodents showing that both acute and chronic exposure to stress during early life or adulthood alter the functional responses in serotonergic neurons [19], reduce the density of 5-HT innervation in the central, basolateral amygdala and the hippocampus [20], increase the density of 5-HT1A receptors in the basolateral amygdala [21], reduce the expression of 5-HT1A and 5-HT1B receptors in the prefrontal cortex [22] and the hippocampus [23,24], increase the expression of the 5-HT transporter, SERT, and the 5-HT synthetizing enzyme, TPH2 in the dorsal raphe nucleus (DRN) [25,26]. Interestingly, comparable neuroplastic changes in brain 5-HT pathways have been observed in alcohol dependence, suggesting that similar mechanisms are involved. ...
Alcohol addiction is a debilitating disorder producing maladaptive changes in the brain, leading drinkers to become more sensitive to stress and anxiety. These changes are key factors contributing to alcohol craving and maintaining a persistent vulnerability to relapse.
Serotonin (5-Hydroxytryptamine, 5-HT) is a monoamine neurotransmitter widely expressed in the central nervous system where it plays an important role in the regulation of mood. The serotonin system has been extensively implicated in the regulation of stress and anxiety, as well as the reinforcing properties of all of the major classes of drugs of abuse, including alcohol. Dysregulation within the 5-HT system has been postulated to underlie the negative mood states associated with alcohol use disorders.
This review will describe the serotonergic (5-HTergic) neuroplastic changes observed in animal models throughout the alcohol addiction cycle, from prenatal to adulthood exposure. The first section will focus on alcohol-induced 5-HTergic neuroadaptations in offspring prenatally exposed to alcohol and the consequences on the regulation of stress/anxiety. The second section will compare alterations in 5-HT signalling induced by acute or chronic alcohol exposure during adulthood and following alcohol withdrawal, highlighting the impact on the regulation of stress/anxiety signalling pathways. The third section will outline 5-HTergic neuroadaptations observed in various genetically-selected ethanol preferring rat lines. Finally, we will discuss the pharmacological manipulation of the 5-HTergic system on ethanol- and anxiety/stress-related behaviours demonstrated by clinical trials, with an emphasis on current and potential treatments.
... Prior studies using immediate early gene mapping have documented the activation of DRN neurons by stress (Paul et al., 2011;. Our previous multilabel studies combining in situ hybridization detection of cell-type specific mRNA markers and immunohistochemical labeling of FOS family proteins, showed that 5-HT neurons represent only a minority (<10%) of DRN cells undergoing FOS-mediated neuroadaptations after repeated exposure to two models of depression (learned helplessness and CSDS) while a comparatively higher proportion of FOS labeled cells expressed GAD mRNA . ...
Regulation of social behaviors is necessary to achieve social inclusion, establish relationships and sustain those relationships through adversity. Impairments in socio-emotional function and competence are prominent and debilitating features of major depression, yet are not traditionally recognized as cardinal symptoms of the disease. However, these deficits often persist in patients whose other mood symptoms have remitted and can predict risk of relapse, indicating an important role as a vulnerability factor. Understanding the neurobiology of socioaffective dysfunction in depression is thus important for determining the pathology of the disorder and developing effective treatments. Human imaging studies of depressive patients have consistently reported abnormal activity in the ventromedial prefrontal cortex (vmPFC), an area important for emotional processing and social cognition. Tracing studies in animals and tractography in humans have shown that the dorsal raphe nucleus (DRN) is a major projection target of the vmPFC. The DRN contains the most serotonin (5-HT) producing neurons in the brain and its output has been shown to regulate behaviors along an affiliative-agonistic axis, however it is neuronally heterogeneous. This thesis investigated the cytoarchitecture of the vmPFC-DRN microcircuit and its relevance to socioaffective behaviors using genetic mapping, whole cell electrophysiology and optogenetics. I showed that GABAergic neurons, which are the primary non-serotonergic neuronal population in the DRN, mediated top-down projections from the vmPFC onto mood-regulating 5-HT neurons and demonstrated the relevance of this pathway in mediating socioaffective decisions using the chronic social defeat stress (CSDS) paradigm. In addition, I used deep brain stimulation of the vmPFC as an antidepressant model to show that therapeutic response may rely on restoring the excitatory/inhibitory balance of inputs to 5-HT neurons. Together, these results will provide a better understanding of socioaffective circuitry and could lead to the development of more effective and efficient strategies to treat mood disorders.
... Moreover, in rats, social defeat by an aggressive male conspecifi c is a natural stressor which is known to induce a state of stress and anxiety as expressed by reduction in behavioral activity, an increase in immobility, and an increase in exploring the enclosed aggressive dominant [ 28 ]. Tuma et al. [ 7 ] tested the hypothesis whether acute or sub-chronic agomelatine would antagonize the negative consequences of a social defeat in male Wistar rats. ...
Anxiety disorders (ADs) are the most common type of psychiatric disorders. Pharmacological options studied for treating ADs may include several compounds such as benzodiazepines, tricyclic antidepressants (TCAs), noradrenergic and specific serotonergic drug (NaSSA), and dual-reuptake inhibitors of serotonin and noradrenaline (SNRIs). However, the selective serotonin reuptake inhibitors (SSRIs) are the gold standard of the treatment of ADs, but some patients do not respond or withdraw the treatment due to adverse effects. Agomelatine, a new antidepressant, acts synergistically on both the melatonergic and the 5-HT2c receptors and has been shown effective in the treatment of major depression. Moreover, there is evidence that suggests efficacy of agomelatine in the treatment of ADs. Therefore, the aim of this chapter is to review the current literature about agomelatine in the treatment of ADs. The clinical trials evaluating agomelatine in the treatment of generalized anxiety disorder are few but, overall, encouraging on establishing its efficacy. Apart from some interesting case reports, to date, no large studies are present in literature about agomelatine and other ADs such as panic disorder, social anxiety disorder, obsessive-compulsive disorder, and post-traumatic stress disorder. Therefore, the clinical efficacy and the relative good tolerability of agomelatine may be further investigated to widen the therapeutic spectrum of ADs.
... This result is consistent with findings by Singh et al. (1990Singh et al. ( , 1994 who reported that repeated systemic dexamethasone injections can restore noisestress induced increases in ex vivo TPH activity in the brain tissue of adrenalectomized rats, whereas adrenalectomy, treatment with a glucocorticoid receptor antagonist, and amygdalar lesions all block ex vivo TPH activity. Activation of c-Fos expression in DRD/DRC serotonergic neurons is observed following exposure to noise stressors (Evans et al., 2009), anxiogenic stimuli such as exposure to an open-field (Hale et al., 2008), intracerebral injections of CRHrelated peptides including the CRHR2-selective agonist urocortin 2 (Evans et al., 2009), systemic treatment with various anxiogenic drugs (Abrams et al., 2005), inescapable shock (Rozeske et al., 2011), and social defeat (Paul et al., 2011). Collateral projections from the DRD/DRC innervate functionally related targets that are in control of anxiety-like behavior and stress responses . ...
Integrating independent but converging lines of research on brain function and neurodevelopment across scales, this article proposes that serotonin 2A receptor (5-HT2AR) signaling is an evolutionary and developmental driver and potent modulator of the macroscale functional organization of the human cerebral cortex. A wealth of evidence indicates that the anatomical and functional organization of the cortex follows a unimodal-to-transmodal gradient. Situated at the apex of this processing hierarchy - where it plays a central role in the integrative processes underpinning complex, human-defining cognition - the transmodal cortex has disproportionately expanded across human development and evolution. Notably, the adult human transmodal cortex is especially rich in 5-HT2AR expression, and recent evidence suggests that, during early brain development, 5-HT2AR signaling on neural progenitor cells stimulates their proliferation - a critical process for evolutionarily-relevant cortical expansion. Drawing on multimodal neuroimaging and cross-species investigations, we argue that, by contributing to the expansion of the human cortex, and being prevalent at the apex of its hierarchy in the adult brain, 5-HT2AR signaling plays a major role in both human cortical expansion and functioning. Due to its unique excitatory and downstream cellular effects, neuronal 5-HT2AR agonism promotes neuroplasticity, learning, and cognitive and psychological flexibility in a context-(hyper)sensitive manner with therapeutic potential. Overall, we delineate a dual role of 5-HT2ARs in enabling both the expansion and modulation of the human transmodal cortex.
Behaviors associated with distress can affect the anxiety-like states in observers and this social transfer of affect shapes social interactions among stressed individuals. We hypothesized that social reactions to stressed individuals engage the serotonergic dorsal raphe nucleus (DRN) which promotes anxiety-like behavior via postsynaptic action of serotonin at serotonin 2C (5-HT2C) receptors in the forebrain. First, we inhibited the DRN by administering an agonist (8-OH-DPAT, 1 μg in 0.5 μL) for the inhibitory 5-HT1A autoreceptors which silences 5-HT neuronal activity. 8-OH-DPAT prevented the approach and avoidance, respectively, of stressed juvenile (PN30) or stressed adult (PN60) conspecifics in the social affective preference (SAP) test in rats. Similarly, systemic administration of a 5-HT2C receptor antagonist (SB242084, 1 mg/kg, i.p.) prevented approach and avoidance of stressed juvenile or adult conspecifics, respectively. Seeking a locus of 5-HT2C action, we considered the posterior insular cortex which is critical for social affective behaviors and rich with 5-HT2C receptors. SB242084 administered directly into the insular cortex (5 μM in 0.5 μL bilaterally) interfered with the typical approach and avoidance behaviors observed in the SAP test. Finally, using fluorescent in situ hybridization, we found that 5-HT2C receptor mRNA (htr2c) is primarily colocalized with mRNA associated with excitatory glutamatergic neurons (vglut1) in the posterior insula. Importantly, the results of these treatments were the same in male and female rats. These data suggest that interactions with stressed others require the serotonergic DRN and that serotonin modulates social affective decision-making via action at insular 5-HT2C receptors.
Social stressors negatively impact social function, and this is mediated by the amygdala across species. Social defeat stress is an ethologically relevant social stressor in adult male rats that increases social avoidance, anhedonia, and anxiety-like behaviors. While amygdala manipulations can mitigate the negative effects of social stressors, the impact of social defeat on the basomedial subregion of the amygdala is relatively unclear. Understanding the role of the basomedial amygdala may be especially important, as prior work has demonstrated that it drives physiological responses to stress, including heart-rate related responses to social novelty. In the present study, we quantified the impact of social defeat on social behavior and basomedial amygdala neuronal responses using anesthetized in vivo extracellular electrophysiology. Socially defeated rats displayed increased social avoidance behavior towards novel Sprague Dawley conspecifics and reduced time initiating social interactions relative to controls. This effect was most pronounced in rats that displayed defensive, boxing behavior during social defeat sessions. We next found that socially defeated rats showed lower overall basomedial amygdala firing and altered the distribution of neuronal responses relative to the control condition. We separated neurons into low and high Hz firing groups, and neuronal firing was reduced in both low and high Hz groups but in a slightly different manner. This work demonstrates that basomedial amygdala activity is sensitive to social stress, displaying a distinct pattern of social stress-driven activity relative to other amygdala subregions.
Social interactions play an important role in our daily lives and can profoundly impact our health for better and worse. To better understand the neural circuitry underlying social behavior, we focused on neural circuits involving vasopressin neurons of the bed nucleus of the stria terminalis (BNST) and serotonin neurons of the dorsal raphe (DR). Previous research shows that BNST vasopressin neurons are activated in male mice by interaction with a female and that vasopressin indirectly excites serotonin neurons. In our studies, we tested the hypothesis that specific social interactions would also activate neurons in the DR, specifically vasopressin 1A receptor (Avpr1a)-expressing neurons, which may be direct targets of the BNST vasopressin neurons. Using in separate experiments immunohistochemistry and in situ hybridization, we found that male and female subjects exposed to a female conspecific show activation in the DR, and the activated neurons include populations of Avpr1a-expressing and other non-serotonergic, non-Avpr1a neurons in roughly equal numbers. Avpr1a neurons in the DR constitute a largely undocumented neuron population. Electrophysiological data suggest that most DR Avpr1a neurons behave like fast spiking interneurons found in other brain regions. Examination of RNAseq and in situ hybridization data suggests that there are glutamatergic, GABAergic, and serotonergic subtypes of Avpr1a neurons in the DR. Together our data support a model in which a subset of vasopressin-responsive interneurons in the DR may relay stimulus specific social signals from the forebrain BNST to the serotonergic DR system, which could help direct prosocial stimulus specific behavioral responses.
The dorsal raphe nucleus (DRN) participates in stress responses and in mood regulation via its ascending release of serotonin (5-HT) onto neural circuits within the forebrain. Although the 5-HT DRN region is easily defined via 5-HT-expressing DRN neurons, the neuroarchitecture and microcircuitry that confer its multifunctionality have remained incompletely understood and have required further investigation. In this present study, neurochemical interactions within different subregions of the rat DRN were precisely analyzed. We found that 97.5% of GABAergic neurons in the DRN expressed ionotropic 5-HT3A receptors (5-HT3ARs), whereas there were only rare parvalbumin (PV)-positive or somatostatin (SOM)-positive GABAergic neurons. Furthermore, corticosterone administration into male rats as a rodent model of depression induced significantly higher c-Fos expression in 5-HT3AR-positive GABAergic neurons compared to that in 5-HT neurons within the DRN. Taken together, our findings suggest that 5-HT3AR-positive GABAergic neurons in the DRN participate in responses to stress hormones in a rat model of depression.
Transport stress causes not only physiological changes but also behavioral responses, including anxiety-like and depression-like behaviors in animals. The serotonergic system in the brain plays a pivotal role in processing anxiety. This study aimed to explore changes in concentrations of 5-hydroxytryptamine (serotonin), and the expression changes of tryptophan hydroxylase 2 (TPH2) mRNA and protein associated with anxiety-related behavioral responses under transport stress. A model of simulated transport stress was established in 40 adult male Sprague-Dawley rats, including a control group (n = 20) and a transport stress (TS) group (n = 20). The results showed that the rats in the TS group exhibited an increased feeding latency in the novelty-suppressed feeding test and a reduced frequency and dwelling time in the central area in the open-field test (OFT). Two hours following the final behavioral test, blood samples were collected. Creatine kinase (CK) activities and glucose and corticosterone concentrations in serum were significantly higher in the rats in the TS group than in the control group. Transport stress also significantly reduced the concentrations of 5-hydroxytryptamine in the hippocampus, striatum, and raphe nuclei and also reduced the expression levels of mRNA and protein for TPH2 in the raphe nuclei. Notably, the number of Fos-immunoreactive neurons was higher in the dorsal raphe nucleus under transport stress, whereas the number of 5-hydroxytryptamine-positive neurons was significantly lower. These findings are consistent with the hypothesis that the 5-hydroxytryptamine transmitter in the hippocampus, striatum, and raphe nuclei is involved in processing anxiety-related behavioral responses under transport stress.
• Lay summary
• Physiological and psychological stress responses were induced in a rat model of simulated transport stress. We examined whether serotonin in the brain may be involved in mediating behavioral responses following exposure to transport stress. Tissue concentrations of serotonin in rat brain regions, including the hippocampus, striatum, and raphe nuclei, were reduced following exposure to transport stress. Expression of tryptophan hydroxylase 2 mRNA and protein, which catalyses serotonin synthesis, as well as numbers of serotonin-immunoreactive neurons, were decreased in the brainstem raphe nuclei.
Objective:
Behavioral inhibition plays a key role in animal stress research and developmental research in children. Therefore, we examined the robustness of our multifaceted model of adult social inhibition that comprises behavioral inhibition, interpersonal sensitivity, and social withdrawal components.
Method:
A total of 899 adults completed the 15-item Social Inhibition Questionnaire (SIQ15) and measures of emotional distress. Confirmatory Factor Analysis (CFA), reliability estimates, and correlational and second-order factor analyses were used to examine the robustness of our model.
Results:
CFA (RMSEA = 0.052; NFI = 0.938; CFI = 0.957) and Cronbach's α estimates ≥0.87 confirmed the robustness of our multi-facet social inhibition model based on three correlated inhibition, sensitivity, and withdrawal factors in 560 adults from the general population and in 194 undergraduate students. Inhibition, sensitivity, and withdrawal were stable over time (3-month test-retest correlations ≥ 0.78), and were closely related to the Gest Behavioral Inhibition and PID-5 Withdrawal measures in a clinical sample of 145 cardiac patients. Of note, male cardiac patients reported more inhibition and withdrawal than female patients. Across samples, social inhibition was distinctly different from negative affectivity.
Conclusions:
Our 3-facet model of inhibition, sensitivity and withdrawal was robust across samples, and may promote research on adult social inhibition in population-based and clinical studies.
Kappa opioid receptor activation has been linked to stress and anxiety behavior, thus leading to kappa antagonists being popularized in research as potential anxiolytics. However, while these findings may hold true in standard models, the neuromodulatory effects of social defeat may change the behavioral outcome of kappa opioid receptor activation. Previous research has shown that social defeat can lead to hyperactivity of serotonergic neurons in the dorsal raphe nucleus, and that inhibition of this increase blocks the social deficits caused by defeat. Kappa opioid receptor activation in the dorsal raphe nucleus works to decrease serotonergic activity. We injected the kappa opioid receptor U50,488 directly into the dorsal raphe nucleus of male and female, defeat and control adult California mice. Here we show evidence that U50,488 induces anxiety behavior in control male California mice, but helps relieve it in defeated males. Consistent with previous literature, we find little effect in females adding evidence that there are marked and important sex differences in the kappa opioid system.
Prolonged or repeated exposure to stressful events has been associated with clinical depression in humans, and also produces depressive-like behaviors in rodent models. Depression has been proposed to be associated with reduced reward- motivated learning. Anhedonia is a main symptom of depression, and the concept of anhedonia refers to a reduction of the ability to experience pleasure, as reflected in a diminished interest in rewarding stimuli and pleasurable events. Many studies have suggested that anhedonia could influence life function and increase vulnerability to the development of psychic disease. A possible dysfunction in the reward and motivation systems has been lately proposed to explain the link between anhedonia and depression. It has been hypothesized that a dysregulated reward system may be associated with the development and maintenance of eating disorders. Indeed, anhedonia is considered as a feature of anorexia nervosa and the most commonly co- morbid disorder in patients with eating disorders. Dysfunctions of the hypothalamic-pituitary-adrenal (HPA) axis activity are most commonly found in patients with eating disorders. We have previously reported that rats with stress experiences in early life show depression-like behaviors including anhedonia, binge-like eating when challenged with metabolic or social stressors, and the HPA axis dysfunctions. In this chapter, neural basis of anhedonia associated with stress- induced disordered eating behaviors in animal models will be discussed.
In adult rats, access to a palatable diet can buffer against the effects of stressors. Approximately 10% of all adolescents are repeatedly victimized by their peers raising the possibility that palatable food consumption may be relevant to this developmental window. This study assessed the long-term impact of juvenile social defeat exposure on anxiety and depressive-like behavior and whether daily limited access to a palatable diet moderated these behavioral consequences. We also investigated the impact of the palatable diet on behavior during the defeat sessions. Juvenile rats were exposed to either a different adult resident rat (Stress) or handling (Control) from postnatal day (PD) 28–34. All rats had ad libitum access to either chow alone or both chow and limited access (4 h/day) to palatable food commencing on PD 21. Results showed that during the defeat sessions, juvenile rats with access to the palatable diet spent less time in submissive postures and displayed significantly longer latencies to submit to the resident. In adulthood, previous exposure to juvenile social defeat resulted in a mild anxiogenic profile in the open field among rats with access to Chow only. Furthermore, defeated rats, regardless of diet, displayed reduced locomotor activity and increased social interaction as adults. These findings suggested only minimal enduring negative consequences from juvenile social defeat exposure which made it challenging to assess potential stress-buffering effects of the palatable diet. This was not the case during the defeat sessions where previous exposure to palatable food appeared protective against the acute stressor effects.
Panic disorder (PD) is an anxiety disorder associated with the occurrence of panic attacks, which arise suddenly without warning. Panic disorder represents a serious psychiatric condition and it can induce complications related to the fear of having subsequent panic attacks and avoidance behaviors. Given its importance, many studies have been conducted to elucidate the circuitry involved in this disorder. Clinical and preclinical studies suggest that PD can be modulated by a specific network of brain structures controlling emotional behaviors and autonomic responses. Using animal models that allow measurement of responses related to behavioral and autonomic symptoms of panic attacks in humans, it has been shown that the neuromodulator serotonin plays an inhibitory role in control of panic attacks associated with PD. Understanding the pathways through which serotonergic systems modulate panic-like responses is key to understanding the biological basis of panic attacks and PD, and, consequently, to establishing novel therapeutic strategies for treatment of PD.
There is evidence for sex differences in cocaine addiction from both clinical and preclinical studies. In particular, preclinical studies indicate that females may be more sensitive than males to stress-induced drug seeking. The dorsal hippocampus (DH) is prominently involved in the stress response, as are the locus coeruleus norepinephrine (LC-NE) and dorsal raphe serotonin (DR 5-HT) systems. Moreover, DH receives strong inputs from LC-NE and DR 5-HT neurons. We hypothesized that the stress associated with non-reinforced drug seeking during early abstinence (on extinction day 1 (ED1)) may contribute to drug seeking via ?-adrenergic and 5-HT neurotransmission in DH. We observed decreased drug-seeking behavior on ED1 following 10?mg/kg S-propranolol (?-adrenergic and 5-HT1A/1B receptor antagonist), R-propranolol (5-HT1A/1B receptor antagonist), or racemic propranolol in both male and female rats. ED1 increased Fos expression in DH, LC, and DR, and DH Fos was decreased by systemic S-propranolol. Based on these results, we investigated the effects of blocking 5-HT and ?-adrenoceptor transmission in DH on drug seeking during ED1 by infusing a cocktail of WAY100635 plus GR127935 (5-HT1A/1B receptor antagonists), betaxolol plus ICI-118?551 (?1 and ?2 antagonists), or S-propranolol alone. In males, WAY100635/GR127935 was most effective in reducing drug-seeking on ED1, whereas betaxolol/ICI-118?551 was ineffective. In contrast, S-propranolol was most effective in females in reducing drug seeking on ED1, and WAY100635/GR127935 and betaxolol/ICI-118?551 were each partially effective. Our results indicate that drug seeking during initial abstinence involves 5-HT and ?-adrenergic signaling in female DH, but only 5-HT signaling in male DH.Neuropsychopharmacology advance online publication, 14 September 2016; doi:10.1038/npp.2016.150.
Depression is a heterogeneous disorder characterized by alterations at psychological, behavioural, physiological, neurophysiological, and neurochemical levels. Social stress is a prevalent stress in man, and the repeated social defeat stress model in rats has been proposed as being the rodent equivalent to loss of control, which in subordinate animals produces alterations that resemble several of the cardinal symptoms found in depressed patients. Here, rats followed a resident-intruder protocol for 4 consecutive days during which behavioural, physiological, and electroencephalographic (EEG) parameters were simultaneously monitored in subordinate rats. On day 5, prefrontal dopamine (DA) and hippocampal serotonin (5-HT) as well as corticosterone were measured in submissive rats that had visual, acoustic, and olfactory (but no physical) contact with a dominant, resident conspecific rat. Socially defeated rats demonstrated increases in ultrasonic vocalizations (20-25 KHz), freezing, submissive defensive behaviour, inactivity, and haemodynamic response, while decreases were found in repetitive grooming behaviour and body weight. Additionally, alterations in the sleep-wake architecture were associated with reduced active waking, enhanced light sleep, and increased frequency of transitions from light sleep to quiet wakefulness, indicating sleep instability. Moreover, the attenuation of EEG power over the frequency range of 4.2-30 Hz, associated with a sharp transient increase in delta oscillations, appeared to reflect increased brain activity and metabolism in subordinate animals. These EEG changes were synchronous with a marked increase in body temperature and a decrease in locomotor activity. Furthermore, psychosocial stress consistently increased 5-HT, DA, and corticosterone levels. The increased levels of cortical DA and hippocampal 5-HT during social threat may reflect a coping mechanism to promote alertness and psychological adaptation to provocative and threatening stimuli. These neurophysiological changes are hypothesized to be the consequence of dynamics in monoamine systems, which could be useful markers for disease progression in the aetiology of depression.
A number of studies have indicated a strong correlation between traumatic events during early life and the development of behavioral abnormalities later in life, including psychoemotional disorders such as anxiety and depression. Patients with eating disorders frequently exhibit symptoms of depression and/or anxiety, as well as reporting experiences of childhood abuse, a type of early-life trauma. Dysfunction of the hypothalamic-pituitary-adrenal (HPA) axis is implicated in the pathophysiology not only of anxiety and depression, but also of eating disorders. Neonatal maternal separation and isolation rearing in rodents are well-known animal models of stressful experiences in early life. Many studies have demonstrated their impacts both on the activity of the HPA axis and on the development of psychoemotional disorders later in life. This chapter reviews research using animal models of eating disorders associated with stress in early life. Results suggest that neonatal maternal separation leads to the development of binge-related eating disorders when it is challenged with social or metabolic stressors later in life, in which dysfunctions in the HPA axis and the brain monoaminergic systems may play important roles. Also, social isolation in adolescence induces hyperphagia and depression-like behaviors in female rats, but not in males; a tonic increase of plasma corticosterone seems to be implicated in its underlying mechanism.
In previous studies, we verified that exposure to unpredictable chronic mild stress (UCMS) facilitates avoidance responses in the elevated T-maze (ETM) and increased Fos-immunoreactivity in different brain structures involved in the regulation of anxiety, including the dorsal raphe (DR). Since, it has been shown that the DR is composed of distinct subpopulations of serotonergic and non-serotonergic neurons, the present study investigated the pattern of activation of these different subnuclei of the region in response to this stress protocol. Male Wistar rats were either unstressed or exposed to the UCMS procedure for two weeks and, subsequently, analyzed for Fos-immunoreactivity (Fos-ir) in serotonergic cells of the DR. To verify if the anxiogenic effects observed in the ETM could be generalized to other anxiety models, a group of animals was also tested in the light/dark transition test after UCMS exposure. Results showed that the UCMS procedure decreased the number of transitions and increased the number of stretched attend postures in the model, an anxiogenic effect. UCMS exposure also increased Fos-ir and the number of double-labeled neurons in the mid-rostral subdivision of the dorsal part of the DR and in the mid-caudal region of the lateral wings. In the caudal region of the DR there was a significant increase in the number of Fos-ir. No significant effects were found in the other DR subnuclei. These results corroborate the idea that neurons of specific subnuclei of the DR regulate anxiety responses and are differently activated by chronic stress exposure.
Mice experiencing repeated aggression develop a long-lasting aversion to social contact, which can be normalized by chronic,
but not acute, administration of antidepressant. Using viral-mediated, mesolimbic dopamine pathway–specific knockdown of brain-derived
neurotrophic factor (BDNF), we showed that BDNF is required for the development of this experience-dependent social aversion.
Gene profiling in the nucleus accumbens indicates that local knockdown of BDNF obliterates most of the effects of repeated
aggression on gene expression within this circuit, with similar effects being produced by chronic treatment with antidepressant.
These results establish an essential role for BDNF in mediating long-term neural and behavioral plasticity in response to
aversive social experiences.
Fear-arousing stimuli elicit innate reactions and can reinforce acquisition of new responses. We tested whether mechanisms mediating these conditioned stimulus (CS) properties were isomorphic or dissociable within the amygdala. Rats trained on a fear-conditioning task (CS paired with footshock) were then trained on an escape-from-fear task (EFF) in which the CS reinforced a locomotor response terminating the CS. Lateral nucleus (LA) lesions blocked acquisition of both conditioned freezing responses and the CS's reinforcement of a new response in the EFF task. Central nucleus (CE) lesions blocked conditioned freezing but not the EFF, whereas basal nucleus (B) lesions blocked the EFF but not conditioned freezing. Thus, activation of the LA by a CS seems to trigger conditioned reactions via CE and conditioned aversion via B activation, reduction of which reinforces new actions.
In a series of experiments, the consequences of a single and double social conflict on various behaviours and body weight in rats were studied. Animals were subjected to social defeat by placing them into the territory of an aggressive male conspecific for one hour, either once, or twice at the same time on two consecutive days. To assess the consequences of social defeat, three experiments were performed with independent groups of rats. In the first experiment, an open field test was performed two days after the last conflict. Locomotor activity was strongly reduced after social defeat. There were no differences between the single and double defeat group. To assess the effects of social defeat on subsequent social behaviour, a second experiment was performed in which experimental animals were confronted with an unfamiliar non-aggressive rat two days after a single or double conflict. Social defeat resulted in a reduction of social contact with the unfamiliar conspecific. There was no difference between the single and double conflict group. In the third experiment, the effects of social conflict on food intake, body weight and saccharine preference were measured. Food intake was not affected after a single conflict, but in the double conflict group food intake was decreased for several days. Body weight gain was decreased after both single and double social defeat. The decrease was stronger in the double conflict group. Water intake and saccharine preference were not significantly affected. This study revealed that social defeat in rats causes pronounced changes in various behaviours and body weight. Different aspects of behaviour are differentially affected by defeat with respect to the magnitude and time course of the changes induced. Moreover, different behavioural parameters are differentially sensitive to repetition of the stressor.
Antidepressants are widely used to treat several anxiety disorders, among which generalized anxiety disorder (GAD) and panic disorder (PD). Serotonin (5-HT) is believed to play a key role in the mode of action of these agents, a major question being which pathways and receptor subtypes are involved in each type of anxiety disorder. The dual role of 5-HT in defense hypothesis assumes that 5-HT facilitates defensive responses to potential threat, like inhibitory avoidance, related to anxiety, whereas it inhibits defensive responses to proximal danger, like one-way escape, related to panic. The former action would be exerted at the forebrain, chiefly the amygdala and medial prefrontal cortex (PFC), while the latter would be exerted at the dorsal periaqueductal gray (DPAG) matter of the midbrain. The present review is focused on studies designed to test this hypothesis, performed in animal models of anxiety and panic, as well as in human experimental anxiety tests. The reviewed results suggest that chronic, but not acute, administration of antidepressants suppress panic attacks by increasing the release of 5-HT and enhancing the responsivity of post-synaptic 5-HT1A and 5-HT2A receptors in the DPAG. The attenuation of generalized anxiety, also caused by the same drug treatment, would be due to the desensitization of 5-HT2C receptors and, less certainly, to increased stimulation of 5-HT1A receptors in forebrain structures. This action would result in less activation of the amygdala, medial PFC and insula by warning signals, as shown by the reviewed results obtained with functional neuroimaging in healthy volunteers and patients with anxiety disorders.
Exposure to social stress has been linked to the development and maintenance of mood-related psychopathology; however, the underlying neurobiological changes remain uncertain. In this study, we examined numbers of δFosB-immunoreactive cells in the forebrains of rats subjected to 12 episodes of social defeat. This was achieved using the social conflict model whereby animals are introduced into the home cage of older males ("residents") trained to attack and defeat all such "intruders"; importantly, controls were treated identically except that the resident was absent. Our results indicated that the only region in which δFosB-positive cells were found in significantly higher numbers in intruders than in controls was the infralimbic medial prefrontal cortex (mPFC). This same effect was not apparent using another psychological stressor, noise stress. Cells of the infralimbic mPFC also displayed evidence of chromatin remodeling. We found that exposure to repeated episodes of social defeat increased numbers of cells immunoreactive for histone H3 acetylation, but not for histone H3 phosphoacetylation, in the infralimbic mPFC. Collectively, these findings highlight the importance of the infralimbic mPFC in responding to social stress-a finding that provides insight into the possible neurobiological alterations associated with stress-induced psychiatric illness.
Anxiety disorders, depression and animal models of vulnerability to a depression-like syndrome have been associated with dysregulation of brain serotonergic systems. These effects could result from genetic influences, adverse early life experiences (ELE), or acute stressful life events, all of which can alter serotonergic neurotransmission and have been implicated in determining vulnerability to neuropsychiatric disorders. To evaluate the effects of ELE, adverse experiences during adulthood, and potential interactions between these factors on neuronal tryptophan hydroxylase 2 (tph2) mRNA expression, we investigated in rats the effects of maternal separation (MS)(separation from the dam for 180 min/day from postnatal day 2-14; MS180, a model of vulnerability to a depression-like syndrome), neonatal handling (separation from the dam for 15 min/day from postnatal day 2-14; MS15, a model of decreased stress sensitivity), or normal animal facility rearing (AFR) control conditions, with or without subsequent exposure to adult social defeat, on tph2 mRNA expression in the dorsal raphe nucleus (DR). Among rats exposed to social defeat, MS180 rats had increased tph2 mRNA expression in the DR, while MS15 rats had decreased tph2 mRNA expression compared to AFR rats. Social defeat increased tph2 mRNA expression, but only in MS180 rats and only in the "lateral wings" of the DR, a subdivision of the DR that is part of a sympathomotor command center. Overall, these data demonstrate that ELE and stressful experience during adulthood interact to determine tph2 mRNA expression. These changes in tph2 mRNA expression represent a potential mechanism through which adverse ELEs and stressful life experiences during adulthood may interact to increase vulnerability to stress-related psychiatric disease.
The purpose of the present study was to determine whether lesions of areas projected to by the central amygdaloid nucleus (ACE) would disrupt the classical conditioning of autonomic and/or behavioral emotional responses. The areas studied included 3 projection targets of the ACE: the lateral hypothalamic area (LH), midbrain central gray (CG) region, and bed nucleus of the stria terminalis (BNST). Lesions were made either electrolytically or by microinjection of ibotenic acid, which destroys local neurons without interrupting fibers of passage. Two weeks later, the animals were classically conditioned by pairing an acoustic stimulus with footshock. The next day, conditioned changes in autonomic activity (increases in arterial pressure) and emotional behavior ("freezing," or the arrest of somatomotor activity) evoked by the acoustic conditioned stimulus (CS) were measured during extinction trials. Electrolytic and ibotenic acid lesions of the LH interfered with the conditioned arterial pressure response, but did not affect conditioned freezing. Electrolytic lesions of the rostral CG disrupted conditioned freezing but not conditioned changes in arterial pressure. Ibotenic acid injected into the rostral CG reduced neither the arterial pressure nor the freezing response. Injection of ibotenic acid in the caudal CG, like electrolytic lesions of the rostral CG, disrupted the freezing, but not the arterial pressure response. Injection of ibotenic acid into the BNST had no effect on either response. These data demonstrate that neurons in the LH are involved in the autonomic, but not the behavioral, conditioned response pathway, whereas neurons in the caudal CG are involved in the behavioral, but not the autonomic, pathway. Different efferent projections of the central amygdala thus appear to mediate the behavioral and autonomic concomitants of conditioned fear.
During an agonistic encounter, subordinate male hamsters display defensive and submissive postures and show increased secretion of glucocorticoids, whereas dominant males do not. To determine whether specific neuronal pathways are activated during the behavioral and neuroendocrine responses of subordinate males, expression of c-fos mRNA within the brains of subordinate males was compared with the pattern in dominant males after fighting. After 1 week of handling, pairs of hamsters were either swapped between cages (handled control males), or were allowed to interact for 30 min [dominant (DOM) males and subordinate (SUB) males]. A second group of control animals that received no handling or social stimulation (unhandled control males) were also included. After testing, all animals were killed by decapitation, their brains were removed for c-fos in situ hybridization, and trunk blood was collected for analysis of plasma cortisol and corticosterone levels. Exposure of males to their partner's cage for 30 min resulted in increased expression of c-fos mRNA in multiple brain regions. In addition, fighting increased c-fos expression in the medial amygdaloid nucleus of both DOM and SUB males as well as having more selective effects. In DOM males, c-fos expression was elevated within the supraoptic nucleus of the hypothalamus. In SUB males, c-fos expression increased within a multitude of brain areas, including cingulate cortex, lateral septum, bed nucleus of the stria terminalis, medial preoptic area, several hypothalamic nuclei, central amygdaloid nucleus, amygdalohippocampal area, dorsal periaqueductal gray, dorsal raphe, cuneiform nucleus, and locus coeruleus. These findings are discussed in relation to neurocircuits associated with behavioral arousal and stress.
The dorsal raphe nucleus through its extensive efferents has been implicated in a great variety of physiological and behavioural functions. However, little is know about its afferents. Therefore, to identify the systems likely to influence the activity of serotonergic neurons of the dorsal raphe nucleus, we re-examined the forebrain afferents to the dorsal raphe nucleus using cholera toxin b subunit and Phaseolus vulgaris-leucoagglutinin as retrograde or anterograde tracers. With small cholera toxin b subunit injection sites, we further determined the specific afferents to the ventral and dorsal parts of the central dorsal raphe nucleus, the rostral dorsal raphe nucleus and the lateral wings. In agreement with previous studies, we observed a large number of retrogradely-labelled cells in the lateral habenula following injections in all subdivisions of the dorsal raphe nucleus. In addition, depending on the subdivision of the dorsal raphe nucleus injected, we observed a small to large number of retrogradely-labelled cells in the orbital, cingulate, infralimbic, dorsal peduncular, and insular cortice, a moderate or substantial number in the ventral pallidum and a small to substantial number in the claustrum. In addition, we observed a substantial to large number of cells in the medial and lateral preoptic areas and the medial preoptic nucleus after cholera toxin b subunit injections in the dorsal raphe nucleus excepting for those located in the ventral part of the central dorsal raphe nucleus, after which we found a moderate number of retrogradely-labelled cells. Following cholera toxin b subunit injections in the dorsal part of the central dorsal raphe nucleus, a large number of retrogradely-labelled cells was seen in the lateral, ventral and medial parts of the bed nucleus of the stria terminalis whereas only a small to moderate number was visualized after injections in the other dorsal raphe nucleus subdivisions. In addition, respectively, a substantial and a moderate number of retrogradely-labelled cells was distributed in the zona incerta and the subincertal nucleus following all tracer injections in the dorsal raphe nucleus. A large number of retrogradely-labelled cells was also visualized in the lateral, dorsal and posterior hypothalamic areas and the perifornical nucleus after cholera toxin b subunit injections in the dorsal part of the central raphe nucleus and to a lesser extent following injections in the other subdivisions. We further observed a substantial to large number of retrogradely-labelled cells in the tuber cinereum and the medial tuberal nucleus following cholera toxin b subunit injections in the dorsal part of the central dorsal raphe nucleus or the lateral wings and a small to moderate number after injections in the two other dorsal raphe nucleus subdivisions. A moderate or substantial number of labelled cells was also seen in the ventromedial hypothalamic area and the arcuate nucleus following cholera toxin injections in the dorsal part of the central dorsal raphe nucleus and the lateral wings and an occasional or small number with injection sites located in the other subdivisions. Finally, we observed, respectively, a moderate and a substantial number of retrogradely-labelled cells in the central nucleus of the amygdala following tracer injections in the ventral or dorsal parts of the central dorsal raphe nucleus and a small number after injections in the other subnuclei. In agreement with these retrograde data, we visualized anterogradely-labelled fibres heterogeneously distributed in the dorsal raphe nucleus following Phaseolus vulgaris-leucoagglutinin injections in the lateral orbital or infralimbic cortice, the lateral preoptic area, the perifornical nucleus, the lateral or posterior hypothalamic areas, the zona incerta, the subincertal nucleus or the medial tuberal nucleus. (ABSTRACT TRUNCATED)
Previously-reported evidence showed that freezing to a context previously associated with footshock is impaired by lesion of the ventral periaqueductal gray (vPAG). It has also been shown that stepwise increase in the intensity of the electrical stimulation of the dorsal periaqueductal gray (dPAG) produces alertness, then freezing, and finally escape. These aversive responses are mimicked by microinjections of GABA receptor antagonists, such as bicuculline, or blockers of the glutamic acid decarboxylase (GAD), such as semicarbazide, into the dPAG. In this work, we examined whether the expression of these defensive responses could be the result of activation of ventral portion of the periaqueductal gray. Sham- or vPAG electrolytic-lesioned rats were implanted with an electrode in the dPAG for the determination of the thresholds of freezing and escape responses. The vPAG electrolytic lesions were behaviorally verified through a context-conditioned fear paradigm. Results indicated that lesion of the vPAG disrupted conditioned freezing response to contextual cues associated with footshocks but did not change the dPAG electrical stimulation for freezing and escape responses. In a second experiment, lesion of the vPAG also did not change the amount of freezing and escape behavior produced by microinjections of semicarbazide into the dPAG. These findings indicate that freezing and escape defensive responses induced by dPAG stimulation do not depend on the integrity of the vPAG. A discussion on different neural circuitries that might underlie different inhibitory and active defensive behavioral patterns that animals display during threatening situations is presented.
Social defeat is an important event in the life of many animals, and forms part of the process of social control. Adapting to social defeat is thus an intrinsic part of social "homeostasis", and mal-adaptation may have pathological sequelae. Experimental models of social defeat (e.g. inter-male aggression) have existed for many years. However, very few studies have investigated the changes in brain activity in male animals exposed to the social stress of being defeated by another conspecific male, and in all these studies the expression of the immediate-early gene c-fos has been used as the marker of neuronal activity. In general, the results obtained inform that many areas of the brain, especially those involved in the general stress response, increase their activity when animals are exposed to an acute defeat. However, when animals are defeated repeatedly over many consecutive days, the level of activation of the brain shows different patterns of adaptation depending on the brain areas (varying from complete habituation to persistent activation). Discrepancies between studies may be due to differences in the experimental procedure. On the other hand, further research has to be conducted in order to understand what these changes in the brain activity mean in relation to the other stress responses to social defeat. Furthermore, knowing that the corresponding protein products of many immediate-early genes are transcription factors that can promote or inhibit the expression of target genes, research following this approach is also necessary.
The role of serotonergic systems in regulation of behavioral arousal and sleep-wake cycles is complex and may depend on both the receptor subtype and brain region involved. Increasing evidence points toward the existence of multiple topographically organized subpopulations of serotonergic neurons that receive unique afferent connections, give rise to unique patterns of projections to forebrain systems, and have unique functional properties. A better understanding of the properties of these subpopulations of serotonergic neurons may aid in the understanding of the role of serotonergic systems in regulation of behavioral arousal, sleep-wake cycles and other physiological and behavioral responses attributed to serotonin. In this chapter, we outline evidence for multiple serotonergic systems within the midbrain and pontine raphe complex that can be defined based on cytoarchitectonic and hodological properties. In addition, we describe how these topographically organized groups of serotonergic neurons correspond to the six major ascending serotonergic tracts innervating the forebrain.
Procedures are given in the report for determining statistically whether the highest observation, or the lowest observation, or the highest and lowest observations, or the two highest observations, or the two lowest observations, or perhaps more of the observations in the sample may be considered to be outlying observations or discrepant values. Statistical tests of significance are useful in this connection either in the absence of assignable physical causes or to support a practical judgement that some of the experimental observations are aberrant. Both the statistical formulae and illustrative applications of the procedures to practical examples are given, thus representing a rather complete treatment of significance tests for outliers in single univariate samples.
The ability of 5-HT1 receptor agonists to modulate a chemically induced defence response has been studied in Lister hooded rats. Microinjections of the excitatory amino acidd,l-homocysteic acid (DLH) in both rostral and caudal dorsal periaqueductal gray matter (PAG) caused explosive motor behaviour characteristic of defence. This behaviour was quantified in terms of response duration, arena revolutions and number of defensive jumps. Direct administration into the PAG of either 5-carboxamidotryptamine (5-CT) or 8-hydroxy-2-(di-n-propylamino) tetralin (8-OHDPAT) produced behaviours (decreased exploratory rearing, dose related onset of flat body posture) indicative of 5-HT1A receptor activation. Pretreatment with either 5-CT or 8-OHDPAT directly in the PAG caused a significant attenuation, and in some cases a complete abolition, of the DLH evoked response. These agonists share high affinity in vitro for the 5-HT1A receptor. Thus the results suggest that in vivo activation of 5-HT1A receptors mediates an antiaversive reponse with respect to defensive behaviour elicited by specific chemical stimulation of the dorsal PAG.
Anxious states are emotional states associated with an increase in avoidance behavior, particularly in situations where a conflict between approach and avoidance occurs, as when both potential rewarding outcomes and potential aversive outcomes exist. An important component of anxious states is that they are trans-situational, transferring from one situation to another. Serotonergic systems play an important role in modulating anxious states. Serotonergic signaling can either facilitate or attenuate anxious states, depending on the site of action and the specific serotonin receptor subtype involved. In this chapter, we present an operational definition of anxious states, describe a number of stimuli that can induce anxious states with distinct time-courses, and consider the evidence that serotonergic systems play a role in modulation of these anxious states. We focus on a subpopulation of serotonergic neurons that projects to forebrain limbic structures implicated in the regulation of anxious states, including the ventral hippocampus/subiculum and the basolateral amygdaloid complex. We then compare and contrast this serotonergic system with another serotonergic system implicated in the inhibition of panic-like responses, a principal component of a subset of anxiety disorders in humans.
Dysfunction of serotonergic systems is thought to play an important role in a number of neurological and psychiatric disorders. Recent studies suggest that there is anatomical and functional diversity among serotonergic systems innervating forebrain systems involved in the control of physiologic and behavioral responses, including the control of emotional states.
Here, we highlight the methods that have been used to investigate the heterogeneity of serotonergic systems and review the evidence for the unique anatomical, hodological, and functional properties of topographically organized subpopulations of serotonergic neurons in the midbrain and pontine raphe complex.
The emerging understanding of the topographically organized synaptic regulation of brainstem serotonergic systems, the topography of the efferent projections of these systems, and their functional properties, should enable identification of novel therapeutic approaches to treatment of neurological and psychiatric conditions that are associated with dysregulation of serotonergic systems.
Social stress has been linked to several neuropsychiatric diseases, including depression, which is a debilitating disease that has genetic, environmental, and epigenetic underpinnings.
This study examined the effects of repeated social defeat on both depressive-like behaviors and histone acetylation in the hippocampus, amygdala, and dorsal prefrontal cortex of male Sprague-Dawley rats.
Subjects were exposed to four consecutive social defeats. Depressive-like behaviors were assayed in the sucrose preference, forced swim, contextual fear, and social approach and avoidance tests. Histone H3 and H4 acetylation in the hippocampus, amygdala, and prefrontal cortex were examined by Western blots under basal conditions and at several time points. We also investigated the potential involvement of N-methyl-D: -aspartic acid (NMDA) receptors and glucocorticoid receptors (GR) by injecting respective antagonists prior to each social defeat and examining their effect on histone acetylation in the hippocampus.
Social defeat resulted in behavioral changes in the forced swim, social avoidance, and contextual fear tests nearly 6 weeks after defeat, with no change in sucrose preference. Additionally, histone H3 acetylation was increased in the hippocampus 30 min following the last defeat and was not blocked by antagonism of either NMDA or GR receptors. There were no changes in histone H4 acetylation.
These results indicate that social defeat induces several long-lasting depressive-like behaviors in rats and induces a significant, short-lived increase in H3 acetylation in the hippocampus, although the underlying mechanism behind this change warrants further investigation.
We examined the roles of the amygdala and hippocampus in the formation of emotionally relevant memories using an ethological model of conditioned fear termed conditioned defeat (CD). Temporary inactivation of the ventral, but not dorsal hippocampus (VH, DH, respectively) using muscimol disrupted the acquisition of CD, whereas pretraining VH infusions of anisomycin, a protein synthesis inhibitor, failed to block CD. To test for a functional connection between the VH and basolateral amygdala (BLA), we used a classic functional connectivity design wherein injections are made unilaterally in brain areas either on the same or opposite sides of the brain. A functional connection between the BLA and VH necessary for the acquisition of CD could not be found because unilateral inactivation of either BLA alone (but not either VH alone) was sufficient to disrupt CD. This finding suggested instead that there may be a critical functional connection between the left and right BLA. In our final experiment, we infused muscimol unilaterally in the BLA and assessed Fos immunoreactivity on the contralateral side following exposure to social defeat. Inactivation of either BLA significantly reduced defeat-induced Fos immunoreactivity in the contralateral BLA. These experiments demonstrate for the first time that whereas the VH is necessary for the acquisition of CD, it does not appear to mediate the plastic changes underlying CD. There also appears to be a critical interaction between the two BLAs such that bilateral activation of this brain area must occur in order to support fear learning in this model, a finding that is unprecedented to date.
Experience with behavioral control over tailshock (escapable shock, ES) has been shown to block the behavioral and neurochemical changes produced by later uncontrollable tail shock (inescapable shock, IS). The present experiments tested, in rats, whether the protective effect of control over tailshock extends beyond reducing the behavioral and neurochemical impact of a subsequent tailshock experience to stressors that are quite different. Social defeat (SD) was chosen as the second stress experience because it has few if any cues in common with tailshock. SD produced shuttlebox escape learning deficits ("learned helplessness") and reduced juvenile social investigation 24 h later, as does IS. IS is notable for inducing a large increase in dorsal raphe nucleus (DRN) serotonergic (5-HT) activity as measured by extracellular levels of 5-HT within the DRN, and SD did so as well. ES occurring 7 days before SD blocked this SD-induced DRN activation, as well as the SD-induced interference with shuttlebox escape and reduction in social investigation. Prior exposure to yoked IS did not reduce the DRN 5-HT activation or later behavioral effects produced by SD, and thus the proactive stress-blunting effects of ES can be attributed to it's controllability. Thus, ES confers a very general protection to the impact of a subsequent stress experience.
Serotonergic (5-HT) neurons in the dorsal raphe nucleus (DRN) have been implicated in stress-induced changes in behavior. Previous research indicates that stressful stimuli activate 5-HT neurons in select subregions of the DRN. Uncontrollable stress is thought to sensitize 5-HT neurons in the DRN and allow for an exaggerated 5-HT response to future stimuli. In the current study, we tested the hypothesis that following aggressive encounters, losing male Syrian hamsters would exhibit increased c-Fos immunoreactivity in 5-HT DRN neurons compared to winners or controls. In addition, we tested the hypothesis that losers would have decreased 5-HT1A mRNA levels in the DRN compared to winners or controls. We found that a single 15-min aggressive encounter increased c-Fos expression in 5-HT and non-5-HT neurons in losers compared to winners and controls. The increased c-Fos expression in losers was restricted to ventral regions of the rostral DRN. We also found that four 5-min aggressive encounters reduced total 5-HT1A mRNA levels in the DRN in losers compared to winners and controls, and that differences in mRNA levels were not restricted to specific DRN subregions. These results suggest that social defeat activates neurons in select subregions of the DRN and reduces message for DRN 5-HT1A autoreceptors. Our results support the hypothesis that social stress can activate 5-HT neurons in the DRN, reduce 5-HT1A autoreceptor-mediated inhibition, and lead to hyperactivity of 5-HT neurons.
While reward-dependent facilitation of phasic dopamine signaling is well documented at both the cell bodies and terminals, little is known regarding fast dopamine transmission under aversive conditions. Exposure to aggressive confrontation is extremely aversive and stressful for many species including rats. The present study used fast-scan cyclic voltammetry and multiunit recording to determine if aggressive encounters and subsequent social defeat affect burst firing of ventral tegmental area (VTA) dopamine neurons and accumbal dopamine transients in defeated rats. Significant increases in the frequency of transient dopamine release were observed during interactions with an aggressive rat but not with a familiar cage mate. In agreement with voltammetric results, significant increases in burst frequency were detected in the VTA dopamine firing patterns during an aggressive confrontation; however, the number of spikes per burst remained unchanged. We found that neurons with lower burst rates under home cage conditions did not switch from nonbursting to bursting types, while neurons with higher burst levels showed amplified increases in bursting. This study demonstrates for the first time that aggressive confrontations in defeated rats are associated with increases in phasic dopamine transmission in the mesolimbic pathway.
Depressed suicide patients have elevated expression of neuronal tryptophan hydroxylase 2 (TPH2) mRNA and protein in midbrain serotonergic neurons, as well as increases in brain serotonin turnover. The mechanisms underlying these changes are uncertain, but increased TPH2 expression and serotonin turnover could result from genetic influences, adverse early life experiences, or acute stressful life events, all of which can alter serotonergic neurotransmission and have been implicated in determining vulnerability to major depression. Emerging evidence suggests that there are several different stress-related subsets of serotonergic neurons, each with a unique role in the integrated stress response. Here we review our current understanding of how genetic and environmental factors may influence TPH2 mRNA expression and serotonergic neurotransmission, focusing in particular on the dorsomedial part of the dorsal raphe nucleus. This subdivision of the dorsal raphe nucleus is selectively innervated by key forebrain structures implicated in regulation of anxiety states, it gives rise to projections to a distributed neural system mediating anxiety states, and serotonergic neurons within this subdivision are selectively activated by a number of stress- and anxiety-related stimuli. A better understanding of the anatomical and functional properties of specific stress- or anxiety-related serotonergic systems should aid our understanding of the neural mechanisms underlying the etiology of anxiety and affective disorders.
When Syrian hamsters (Mesocricetus auratus) are defeated by a larger, more aggressive opponent, they subsequently produce more defensive and submissive behaviors and less chemosensory investigation and aggression, even when they are paired with a smaller, non-aggressive intruder. This persistent change in the behavior of defeated animals has been termed conditioned defeat. In the present study, we tested the hypothesis that the bed nucleus of the stria terminalis (BNST) is important for the acquisition and expression of conditioned defeat. We found that the GABA(A) receptor agonist muscimol infused into the BNST immediately prior to initial defeat training failed to disrupt the acquisition of conditioned defeat, while muscimol infused prior to testing caused a significant reduction in submissive/defensive behaviors and an increase in investigatory behaviors of the non-aggressive intruder. These results indicate that (1) the BNST, unlike the amygdala, does not appear to be critically involved in the consolidation process related to the memory of social defeat and (2) the BNST may be an important site for the execution of fear behaviors associated with social defeat. Considering the high degree of connectivity between the BNST and the amygdala, these findings provide further insight into the neural circuitry governing conditioned defeat and support the view of a functional dissociation between the amygdala and the BNST in the modulation of conditioned fear in an ethologically relevant model.
Social defeat, resulting from the fight for a territory is based on the resident-intruder paradigm. A male rat intruder is placed in the territory of an older, bigger and more aggressive male resident and is defeated. In the present study, a double exposure to social defeat increased sleep fragmentation due to an increased amount of waking and slow-wave-sleep-1 (SWS-1) episodes. Also, social defeat increased the amount of slow-wave-sleep-2 (SWS-2). In repeated exposures to an open field, socially defeated rats showed low central activity and persistent defecation indicating high emotionality. The strongest effects of social defeat on sleep and open field behaviour were seen sub-chronically after stress. Social defeat did not induce changes in rapid eye movement (REM) sleep (e.g. total amount, latency), sleep latency, sexual activity, body weight or adrenal weight. A negative correlation between habituation in open field central activity and total sleep fragmentation indicates a commonality of effects of social defeat on both behaviour and sleep.
In animal models, serotonin (5-HT) activity contributes to stress-induced changes in behavior. Syrian hamsters (Mesocricetus auratus) exhibit a stress-induced change in behavior in which social defeat results in increased submissive and defensive behavior and a complete loss of normal territorial aggression directed toward a novel, non-aggressive opponent. We refer to this defeat-induced change in agonistic behavior as conditioned defeat. In this study we tested the hypothesis that 5-HT activity in the dorsal raphe nucleus (DRN) contributes to the acquisition and expression of conditioned defeat. We investigated whether injection of the selective 5-HT1A agonist flesinoxan (200 ng, 400 ng, or 800 ng in 200 nl saline) into the DRN would reduce the acquisition and expression of conditioned defeat. Additionally, we investigated whether injection of the selective 5-HT1A antagonist WAY 100635 (400 ng in 200 nl saline) into the DRN would enhance the acquisition and expression of conditioned defeat following a sub-optimal social defeat experience. We found that injection of flesinoxan into the DRN before exposure to a 15-min social defeat reduced the amount of submissive and defensive behavior shown at testing. We also found that injection of flesinoxan into the DRN before testing similarly reduced submissive and defensive behavior. In addition, we found that WAY 100635 enhanced conditioned defeat when injected either before social defeat or before testing. These data support the hypothesis that the activity of 5-HT cells in the DRN, as regulated by 5-HT1A autoreceptors, contributes to the formation and display of conditioned defeat. Further, our results suggest that 5-HT release in DRN projection regions augments defeat-induced changes in social behavior.
Striatal extracellular levels of serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were monitored with the microdialysis technique during electrical stimulation of the lateral habenula-dorsal raphe (LHb-NRD) pathway in halothane anaesthetized rats. A new double-loop probe, with an improved recovery factor, was implanted into the head of the caudate-putamen and perfused with Ringer solution containing 1 microM of the 5-HT uptake blocker indalpine. Samples were collected every 15 min and analyzed with HPLC coupled to fluorimetric detection. Low frequency stimulation of the LHb (1.5 and 3 Hz, 0.5 mA) produced no detectable changes in striatal indole levels, whereas 15 Hz stimulation induced a 70% increase in 5-HT release. This effect was most likely mediated by a direct LHb-NRD link, since it persisted after ibotenic acid lesions of the interpeduncular nucleus (which is the major projection area for the medial habenular nucleus), but was completely abolished after transection of the fasciculus retroflexus, which carries the axons of the LHb-NRD pathway. The possible identity of the transmitter operating in the LHb-NRD pathway was investigated by NRD injections of kynurenic acid, a potent blocker of excitatory amino acid transmission, and by NRD injections of the GABA antagonist bicuculline. Kynurenic acid (300 nl, 50 mM) did not by itself induce any detectable changes in spontaneous indole output, but completely blocked the effect of LHb stimulation. Injection of bicuculline (300 nl, 2 mM) increased the striatal 5-HT output by about 70%, and potentiated the effect of LHb stimulation by a further 50%. In none of the experiments performed in this study were there any significant changes in striatal 5-HIAA output. These data are compatible with the idea that excitatory amino acids in the LHb-NRD pathway are involved in the regulation of striatal 5-HT release, and that this influence is modulated by GABAergic synaptic activity at the level of the NRD.
Evolutionary constant serotonin (5-HT) neuronal systems evolved along medial brain structures; yet, wide variations in functionality characterize serotonergic systems in mediating aggressive responses in species ranging from lobsters, ants, electric fish, and rodents to primates. So far, the attempts to correlate cerebrospinal fluid (CSF) 5-hydroxyindoleacetic acid (5-HIAA) levels with measures of aggression have revealed inverse, direct, or no correlations in different nonhuman primate species. It is difficult to harmonize the occasional correlations between CSF 5-HIAA and adaptive aggressive acts in nonhuman primates (a) with clinically diagnosed suicidal or impulsive individuals, and (b) with the biochemical, anatomical, and presumably functional differentiation of 5-HT pathways and receptor subtypes. Eltoprazine, a mixed 5-HT1A/B agonist, and meta-trifluoro-methylphenyl-piperazine HCl (TFMPP), a more selective 5-HT1B agonist, specifically decrease aggressive behavior in several animal species and situations in both sexes without detriment to other social, exploratory, or motoric activities. A definite role for 5-HT1A, 5-HT2, and 5-HT3 receptor subtypes in the mechanisms mediating aggressive behaviors has to await the development of selective agonists and antagonists, respectively.
Fos protein can trans-activate AP-1-dependent gene expression and trans-repress the c-fos promoter. Although we find that trans-repression is enhanced by coexpression of c-Jun, it does not require any of the AP-1 or ATF sites in the mouse c-fos promoter. A major target for repression is the serum response element (SRE). Fos mutants with an impaired leucine zipper are defective in trans-repression and transformation, suggesting that these functions involve the formation of Fos protein complexes. In contrast, mutations that abolish DNA binding of Fos enhance trans-repression but destroy the transforming potential of Fos. In addition, v-Fos protein efficiently transforms but is unable to trans-repress. These findings point to different mechanisms involved in trans-activation and trans-repression and suggest that trans-repression of the type described here is neither sufficient nor required for Fos-induced transformation.
Serum-induced transcription of the proto-oncogene fos is under negative feedback regulation mediated by the fos protein. The fos promoter region responsive to repression is also required for serum inducibility and binds a nucleoprotein complex in which the nuclear factor AP-1 is associated with fos protein.
The behavioural and physiological consequences of social status and reciprocal fighting in resident-intruder dyads of Long Evans male rats were evaluated. Before a chronic cohabitation of 10 days, residents and intruders were individually housed for one month to increase their aggressiveness. Control animals included isolates, i.e., animals kept individually housed throughout the experiment and pair-housed rats, i.e., pairs of rats housed together from their rats in the laboratory. In 19 out of 20 dyads, a clear dominance relationship developed with an advantage to the resident in 68% of the cases. Dominants showed more exploratory activity than subordinates in a open-field test at the end of the cohabitation period; subordinates groomed longer than animals from other experimental groups. Dominants had lower pain thresholds than individually and pair-housed animals. Both dominants and subordinates had higher tyrosine hydroxylase enzymatic activities in the left adrenal than isolated and pair-housed rats. Subordinates lost body weight and had higher plasma corticosteroid concentrations than animals from the other experimental groups. In addition, they had smaller thymus glands and reduced spleen lymphocyte responses to mitogenic stimulation in vitro, in comparison to dominant animals. These results show that subordination in the dyadic resident-intruder paradigm leads to a complex syndrome of behavioural and physiological changes, some of which may be modulated by the intensity of aggressive interactions.
In this study, noradrenergic (NE) terminals in the dorsal raphe were identified by [3H]NE electron microscopic (EM) autoradiography. Lesioning of NE terminals by treatment with the selective catecholamine neurotoxin, 6-hydroxydopamine produced a marked decrease in NE-labelled terminals. [3H]5-HT EM autoradiography of the dorsal raphe produced labelling of cell bodies, dendrites and axons but labelled terminals with synaptic junctions were not observed. Serotonergic (5-HT) neurons were identified at an early stage of degeneration following treatment with the selective 5-HT neurotoxin, 5,7-dihydroxytryptamine (5,7-DHT). When both [3H]NE autoradiography and 5,7-DHT lesioning were combined, a majority of NE-labelled terminals, which formed synaptic specializations, innervated degenerating dendrites. These findings suggest that NE terminals directly innervate 5-HT cells in the dorsal raphe.
In rats treated systemically with either amphetamine, amfonelic acid or apomorphine, large numbers of cells displaying Fos-like immunoreactivity (FLI) could be seen in the lateral zone of the lateral habenula. The induction of FLI by amphetamine could be blocked either by pretreatment with haloperidol or by 6-hydroxydopamine lesions of ascending dopamine fibers at the level of the lateral hypothalamus. In contrast, a variety of stressors selectively induced FLI in the most medial portion of the lateral habenula. These findings support the concept of a functional differentiation of the medial and lateral regions of the lateral habenula and provide further evidence for involvement of the habenula in the circuitry of the basal ganglia.
This experiment was designed to examine the short- and long-term behavioral, cardiovascular, and thermoregulatory responses to brief intermittent agonistic confrontation in rats. The experimental procedure involves resident-intruder confrontations consisting of a 10-min period during which both animals are separated in the home cage of the resident, followed by a brief physical encounter leading to defeat of the intruder and a 10-min period, when the intruder was alone in the home cage of the resident. These 30-min-long confrontations were repeated on 5 consecutive days. Before the first confrontation with a resident, an intruder rat's telemetered heart rate and core temperature show a stable circadian rhythm that is entrained by the light cycle. Acutely, the confrontations produce immediate and large tachycardia and hyperthermia in intruders. A decrease in amplitude of the circadian rhythms for heart rate and core temperature, as detected by cosinor analysis, persists for at least 10 days after the last of five daily brief confrontations with the resident. The defensive upright posture is nearly absent during the first exploration of the resident's home cage, but is displayed by the intruder for one-third of the available time before and after the fifth defeat. Intermittent brief social stress is sufficient to induce profound changes in defensive behavior and long-lasting depression of circadian rhythmicity that persist for weeks.
The effects of the selective 5-HT(1A) agonist, 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OHDPAT) and the selective 5-HT(1A) antagonist, N-[2-[4-(2-methoxyphenyl)-1-piperzinyl]ethyl]-N-(pyridinyl) cyclohexanecarboxamide trichloride (WAY100635) on periaqueductal grey (PAG)-stimulated defence behaviour were tested in the rat. Microinjection of the excitatory amino acid, D, L-homocysteic acid (DLH) into the dorsal region of the PAG produced overt aversive behaviour characteristic of the defence response, consisting of explosive motor behaviours which were quantified in terms of their duration and the number of arena revolutions and jumps made by the animal. Intra-PAG pre-treatment with 8-OHDPAT (3, 10 and 25 nmol in 250 nl) 10 min before DLH stimulation significantly attenuated the defence behaviour. This could be reversed by peripheral application of WAY100635 (0.1 mg/kg). In contrast, peripheral 8-OHDPAT (0.03, 0.1 and 0.3mg/kg) produced a significant potentiation of the DLH response which could also be blocked by peripheral WAY100635. When WAY100635 (10 nmol in 250 nl) alone was given into the PAG a significant increase in DLH induced behaviours was observed whereas peripherally applied WAY100635 (0.1 mg/kg) was without effect. These data support previous findings which indicate that serotonergic modulation of aversive behaviours such as defence can be mediated by 5-HT(1A) receptors. Furthermore there is evidence to indicate a differential involvement of pre- and postsynaptic 5-HT(1A) receptors.
In coronal slices of rat midbrain localised injections of FluoroGold or DiO into the dorsolateral periaqueductal grey matter (PAG) labelled cells retrogradely in the dorsal half of the dorsal raphe nucleus (DRN) and the ipsilateral ventrolateral PAG. Low intensity electrical stimulation in the dorsolateral PAG (22.9 +/- 1.9 microA) evoked antidromic responses in neurones recorded intracellularly in the dorsal subnucleus of the DRN. Antidromic responses could also be evoked in neurones in the ventral DRN and its wings but only at much higher currents (40.9 +/- 2.5 microA) which likely spread to activate axons in the ventrolateral PAG that originated from perikarya in the ventral DRN. The findings are discussed in relation to modulation of the excitability of the aversive system in the dorsolateral PAG by the DRN.
Intraspecific confrontation between male rats represents a biologically relevant form of social stress. C-fos expression has been used to map the pattern of neural activation following either a single (acute) or repeated (10 times) exposure of an intruder male to a larger male in the latter’s home cage. These conditions induce high levels of aggressive interaction.
Sixty minutes after a single defeat, there was intense c-fos expression (quantified using image analysis) in restricted areas of the basal forebrain (including lateral septum, bed nucleus of stria terminalis, lateral preoptic area, lateral hypothalamic area, paraventricular nucleus, and medial and central amygdala) as well as in the autonomic and monoaminergic nuclei of the brainstem (central grey, dorsal and median raphe, locus coeruleus and nucleus of the solitary tract). After the tenth defeat, this pattern was modified despite persistently high levels of aggression. Some areas in the forebrain (bed nucleus of stria terminalis, paraventricular nucleus and medial amygdala) continued to express increased c-fos; others (the septum, lateral hypothalamic area, lateral preoptic area and central amygdala) no longer expressed c-fos. The brainstem response was equally varied: the central grey and the raphe nuclei continued to respond after repeated defeat, whereas the solitary nucleus and locus coeruleus did not. On the other hand, there was no change in the behaviour of intruder rats after repeated defeat.
This study shows the pattern of adaptation at a cellular level in the basal forebrain and brainstem to repeated defeat. As in our previous studies of repeated restraint, modulation in the expression of c-fos following repeated stress is highly regionally specific, suggesting that differential neural processing is involved in adaptation to social stress.
The objective of the present study was to determine how c-fos gene expression in brainstem structures after a brief episode of social defeat stress is related to behavioral sensitization to cocaine challenge. Social stress was defined as defeat in a brief confrontation with an aggressive resident mouse and subsequent 20-min exposure to the resident's threats behind a protective screen. Mice were treated with cocaine (40 mg/kg, i.p.) immediately or 1 week after social defeat stress. Fos-like immunoreactive (Fos-LI) cell nuclei were analyzed in the ventral tegmental area (VTA), dorsal raphe nucleus (DR), periaqueductal grey area (PAG) and locus coeruleus (LC). One episode of social stress induced behavioral sensitization to cocaine as indicated by an augmented locomotor response to a challenge injection 7 days after a single defeat. In naive mice, social stress markedly increased the number of Fos-LI nuclei in the DR, PAG and LC, but not in the VTA. Similarly, cocaine administration resulted in a significantly increased number of Fos-LI nuclei in the same areas. Administration of cocaine immediately following social defeat significantly reduced the number of Fos-LI nuclei in the DR, PAG and LC. Cocaine-induced Fos expression returned in the PAG and DR, but not in the LC, 1 week after social stress. In conclusion, the present results suggest that the presence of brainstem Fos be related to the ability to express stress-induced behavioral sensitization to cocaine.
The dorsal raphe nucleus (DRN) and its serotonergic terminal regions have been suggested to be part of the neural substrate by which exposure to uncontrollable stressors produces poor escape responding and enhanced conditioned fear expression. Such stressor exposure is thought to selectively activate DRN serotonergic neurons in such a way as to render them transiently sensitized to further input. As a result of this sensitized state, behavioral testing procedures are thought to cause excess serotonergic activity in brain regions that control these behaviors. The present studies were conducted to investigate activity in the DRN following exposure to escapable and yoked, inescapable tailshock. Neural activity was characterized using immunohistochemistry to detect the immediate early gene product Fos in serotonin-immunoreactive cells in the DRN. Inescapable tailshock led to greater serotonergic neural activity than did escapable tailshock, supporting the hypothesis that uncontrollable stressors preferentially activate serotonergic neurons in the DRN.
Intraspecific confrontation has been used to study effect of depleting central serotonin on the adaptation of male rats to repeated social stress (social defeat). Four groups of adult male rats were used (serotonin depletion/sham: stressed; serotonin depletion/sham: non-stressed). Central serotonin was reduced (by 59-97%) by a single infusion of the neurotoxin 5,7-dihydroxtryptamine (150 microg) into the cerebral ventricles; levels of dopamine and noradrenaline were unaltered (rats received appropriate uptake blockers prior to neurotoxic infusions). Sham-operated animals received solute only. Rats were then either exposed daily for 10 days to a second larger aggressive male in the latter's home cage, or simply transferred to an empty cage (control procedure). Rats with reduced serotonin failed to show the increased freezing behaviour during the pre-defeat phase of the social interaction test characteristic of sham animals. There was no change in the residents' behaviour. Core temperature increased during aggressive interaction in sham rats, and this did not adapt with repeated stress. By contrast, stress-induced hyperthermia was accentuated in serotonin-reduced rats as the number of defeat sessions increased. Basal core temperature was unaffected by serotonin depletion. Heart rate increased during social defeat, but this did not adapt with repeated stress; serotonin depletion had no effect on this cardiovascular response. Basal corticosterone was increased in serotonin-depleted rats, but the progressive reduction in stress response over days was not altered. C-fos expression in the brain was not altered in control (non-stressed) rats by serotonin reduction in the areas examined, but there was increased expression after repeated social stress in the medial amygdala of 5-HT depleted rats. These experiments show that reduction of serotonin alters responses to repeated social stress in male rats, and suggests a role for serotonin in the adaptive process.
Chronic exposure to stress has been implicated in physical and mental illness, and such experiences can produce alterations in the connectivity and number of neurones within the brain to variations in the expression of specific genes. The purpose of this study was to determine how repeated exposure to social defeat affects neuronal activation patterns within the male Syrian hamster brain. Toward this end, the levels c-fos mRNA were compared among three groups: (1) handled controls (HC); (2) acutely defeated males (AD); and (3) chronically defeated males (15 min aggression daily, 7 days) exposed to an acute challenge (CD). Plasma glucocorticoids were also measured and compared among groups as an index of neuroendocrine activity. The results show a selective pattern of habituation of immediate early gene expression within the brains of chronically defeated males. In particular, c-fos mRNA levels were significantly decreased within the paraventricular nucleus of the hypothalamus (PVN), supraoptic nucleus of the hypothalamus, septohypothalamic nucleus, intermediate subdivision of the lateral septum, central amygdaloid nucleus, and the amygdalohippocampal area in the CD group exposed to an acute challenge when compared to males defeated only once. In contrast, c-fos expression within the anterior and ventromedial nuclei of the hypothalamus, dorsal periaqueductal grey, dorsal raphe, cuneiform nucleus, and locus coeruleus did not differ between AD and CD groups. Similarly, plasma levels of cortisol and corticosterone in CD group were equivalent to those observed after a single defeat experience. We discuss the possibility that decreased expression of c-fos mRNA within the PVN and other brain regions of defeated animals-in the presence of elevated adrenal steroids-may reflect a state of molecular plasticity that could alter neurotransmission within the limbic-hypothalamo-pituitary-adrenocortical axis. In contrast, brain areas that maintain relatively high levels of c-fos mRNA following repeated defeat may reflect processes less likely to adapt such as defensive behaviour.
This paper summarizes the current views on coping styles as a useful concept in understanding individual adaptive capacity and vulnerability to stress-related disease. Studies in feral populations indicate the existence of a proactive and a reactive coping style. These coping styles seem to play a role in the population ecology of the species. Despite domestication, genetic selection and inbreeding, the same coping styles can, to some extent, also be observed in laboratory and farm animals. Coping styles are characterized by consistent behavioral and neuroendocrine characteristics, some of which seem to be causally linked to each other. Evidence is accumulating that the two coping styles might explain a differential vulnerability to stress mediated disease due to the differential adaptive value of the two coping styles and the accompanying neuroendocrine differentiation.
Electrical stimulation of the dorsal periaqueductal gray (DPAG) has been used to induce panic-like behavior in rats. In the present study, we investigated the effect of chronic imipramine treatment on the sensitivity of different 5-HT receptor subtypes in inhibiting aversion induced by electrical stimulation of this brain area. For that, the effects of intra-DPAG administration of the endogenous agonist 5-HT (20 nmol), the 5-HT(1A) receptor agonist 8-OH-DPAT (8 nmol) and the 5-HT(2A/2C) receptor agonist DOI (16 nmol) were measured in female Wistar rats given either chronic injection of imipramine (15 mg/kg, 3 weeks, ip) or saline. The results showed that the three receptor agonists raised the threshold of aversive electrical stimulation in both groups of animals, but this antiaversive effect was significantly higher in rats treated with imipramine. Treatment with imipramine did not change the basal threshold of aversive electrical stimulation measured before intra-DPAG injection of the 5-HT agonists. The results suggest that sensitization of both 5-HT(1A) and 5-HT(2) receptors within the DPAG may be involved in the beneficial effect of imipramine in panic disorder (PD).
Based on WGA-apo-HRP-gold (WG) retrograde tracing, the present study revealed that different subdivisions of the dorsal raphe (DR) such as dorsomedial, ventromedial, lateral wing, and caudal regions receive unique, topographically organized afferent inputs, that are more restricted than previously reported. Phaseolus vulgaris leucoagglutinin anterograde tracing studies confirmed that the medial prefrontal cortex provides the major afferent input to each subdivision of the DR. Double-labeling studies combining WG tracing and glutamate immunostaining indicated that the medial prefrontal cortex, various hypothalamic nuclei including perifornical, lateral, and arcuate nuclei, and several medullary regions such as lateral and medial parabrachial nuclei, and laterodorsal tegmental nucleus provide the major glutamatergic input to each subregion of the DR. It should be noted that the degree of glutamatergic input from these afferent sites was specific for each DR subdivision. The present findings indicated that dorsomedial, ventromedial, lateral wing, and caudal subdivisions of the DR receive excitatory inputs from both cortical and subcortical sites which might be involved in regulation or modulation of a broad range of systems, including sensory and motor functions, arousal and sleep-wake cycle, biorhythmic, cognitive, and affective behaviors.
Here, we describe a biologically relevant model called conditioned defeat that is used to examine behavioral responses to social defeat in Syrian hamsters. In this model experimental animals that are normally aggressive experience social defeat and consequently display high levels of submissive/defensive behavior even in response to non-threatening conspecifics. N-methyl-D-aspartate (NMDA) receptors within the amygdala play an important role in conditioned fear; therefore, the purpose of this study was to examine whether NMDA receptors within the amygdala are necessary for the acquisition and expression of conditioned defeat. Specifically, the present study examined whether bilateral infusions of the NMDA receptor antagonist DL-2-amino-5-phosphonopentanoic acid (AP5; 0.625, 1.25, 2.5, 5.0, 10.0 microg) into the amygdala would block the acquisition of conditioned defeat. Subsequently, we examined whether bilateral infusions of AP5 (0.625, 1.25, 2.5, 5.0 microg) into the amygdala prior to testing would block the expression of conditioned defeat. Infusions of AP5 into the amygdala immediately before the initial social defeat significantly reduced submissive/defensive behavior when hamsters were tested the following day with a non-aggressive intruder. Similarly, infusions of AP5 into the amygdala immediately before exposure to a non-aggressive intruder significantly attenuated the display of submissive/defensive behavior. These data demonstrate that NMDA receptors are necessary for both the acquisition and expression of conditioned defeat. We believe that conditioned defeat is a unique and valuable animal model with which to investigate the neurobiology of fear-related changes in social behavior.
The negative impact of chronic stress at the structure of apical dendrite branches of cornu ammonis 3 (CA3) pyramidal neurons is well established. However, there is no information available on the CA3 dendritic organization related to short-lasting stress, which suffices to produce long-term habituation or sensitization of anxiety behaviors and neuroendocrine responses. Here, we tested the effects evoked by brief stress on the arrangements of CA3 pyramidal neuron dendrites, and the activity-dependent properties of the commissural-associational (C/A) excitatory postsynaptic potentials (EPSPs). Adult male rats were socially defeated followed by 3 weeks without further treatment or as comparison exposed to a regimen of a social defeat every second day for the same time period. We assessed CA3 pyramidal neurons with somatic whole-cell recording and neurobiotin application in acute hippocampal slices. The results from morphometric analysis of post hoc reconstructions demonstrated that CA3 dendrites from repeatedly stressed rats were reduced in surface area and length selectively at the apical cone (70% of control, approximately 280 microm from the soma). Brief stress, however, produced a similar decrease in apical dendritic length (77% of control, approximately 400 microm from the soma), accompanied by an increased length (167% of control) and branch complexity at the basal cone. The structural changes of the dendrites significantly influenced signal propagation by shortening the onset latency of EPSPs and increasing input resistance (r=0.45, P<0.01), of which the first was significantly changed in repeatedly stressed animals. Both brief and repeated stress long-lastingly impaired long-term potentiation of C/A synapses to a similar degree (P<0.05). These data indicate that the geometric plasticity of CA3 dendrites is dissociated from repetition of aversive experiences. A double social conflict suffices to drive a dynamic reorganization, by site-selective elimination and de novo growth of dendrite branches over the course of weeks after the actual experience.