Article

The organization of divergent axonal projections from the midbrain raphe nuclei in the rat

January 1986
The Journal of Comparative Neurology 243(3):363-80

January 1986
243(3):363-80

DOI:10.1002/cne.902430307

Source
PubMed

Authors:

Hisamasa Imai

Juntendo University

Dennis Steindler

University of Florida

The intranuclear organization of divergently projecting neurons of the midbrain raphe in the rat was studied by using double retrograde axonal tracing. Paired injections of the tracers N-[acetyl-3H] WGA and horseradish peroxidase were made within known projection targets of the midbrain raphe (caudate-putamen, amygdala, hippocampus, substantia nigra, and locus coeruleus). After injections of either tracer in the aforementioned targets, retrograde labeled neurons were found mainly ipsilaterally and within midline portions of the dorsal raphe nucleus, its caudal B6 portion, and within the linear and superior central nuclei of the median raphe complex. There are discrete intranuclear distributions of raphe neurons that project to these forebrain and brainstem sites, and there is an overall rostrocaudal topographic order within the raphe with neurons projecting to the neostriatum, amygdala, and substantia nigra residing most rostrally and neurons projecting to the hippocampus and/or locus coeruleus occupying caudal portions of the B6 and superior central nuclei. Such distributions of projection neurons suggest the existence of an “encephalotopic” intranuclear organization within the raphe; that is, each central nervous system structure that receives midbrain raphe projections has its own unique representation within a topographically distinct portion of one or more of the raphe subgroups. These findings suggest an overall functional organization within the midbrain raphe nuclear complex whereby rostral portions are associated with the basal ganglia and related nuclei, and caudal portions relate to the limbic system. An intermediate representation of amygdala-projecting raphe neurons functionally conjoins the two. Collateralized neurons are found within complex zones of overlap in the topographically organized distributions of raphe neurons projecting to functionally related structures.

Integrative physiology of antidepressant drug action

Article

Apr 2012

Previous studies have demonstrated that depressed patients have dysfunction of thermoregulatory cooling, while antidepressants of diverse pharmacological profiles induce sweating as a clinical side effect, suggesting that the pathophysiology of depression as well as antidepressant drug action may involve interactions with thermoregulatory pathways. To test this hypothesis, we investigated interactions between acute whole body warming and treatment with the selective serotonin reuptake inhibitor, citalopram, on body temperature, measured using chronically implanted telemetric recording devices, and antidepressant‐like behavior in the forced swim test in rats. Using a subthreshold dose of citalopram, we found that citalopram, by itself, induced hyperthermia that was comparable to that induced by exposure to increased ambient temperature (37 degrees C) for 85 min. Neither citalopram by itself, nor exposure to increased ambient temperature induced antidepressant‐like effects in the forced swim test. However, when rats were both treated with citalopram and exposed to elevated ambient temperature, rats experienced an exaggerated hyperthermia that was highly correlated with antidepressant‐like behavioral responses. These data provide a rationale for novel therapeutic strategies for the treatment of affective disorders, including development of novel antidepressant drugs.

Molecular guidance of serotonin raphe neurons during development

Thesis

Sep 2016

Teng Teng

In mice, serotonin (5-HT) midbrain neurons are born from embryonic day 10 to 12, and start extending axons, shortly after neurogenesis, both rostrally to the telencephalon and caudally to the brainstem. These projections are highly collateralized but with some degree of topographic organization. In the telencephalon, the pattern of 5-HT innervation arising from the dorsal (B7, B6) or the medial (B5-B8) nuclei differs. However, there are no systematic detailed developmental studies in mice, which are the most extensively used model, in particular for genetic studies. Such data are important to gather in order to analyze the effects of mouse mutations on defined molecular pathway of serotonin neurons. Moreover the guidance molecules that direct these 5-HT raphe neurons to different targets are not known. We performed several studies of 5-HT innervation aimed at detecting how the dorsal and median raphe nuclei are targeted to different forebrain regions during development. We investigated the role of ephrinA-EphA signaling in selective targeting. Our results demonstrate that EphA5 mRNA is selectively expressed in distinct subpopulation of serotonin raphe neurons. Particularly, EphA5 exhibited the highest level in dorsal raphe serotonin neurons (B7). The results of in vitro explant cultures and in vivo electroporation analyses indicated that the ligands of EphA5 (ephrinA5 and ephrinA3) act as repellent factors for the serotonergic axon growth cones. Anterograde tracing in the ephrinA5 -/- mice showed mistargeting of dorsal raphe neurons projections, including the serotonergic projection. Particularly, our analysis of tracing studies show that targeting of the dorsal and median raphe axons to different layers of the olfactory bulb is altered in the ephrinA5 KO. However we do not know at what developmental stage these alterations occur, in particular whether this reflects an alteration in the orientation of ascending fiber tracts, or whether this reflects late developmental maturation when raphe axons collateralize and branch in specific target regions. We have taken advantage a new morphological method, which allows analyzing immunocytochemical labeling in 3_D. 5-HT immunolabeling, in whole brain serotonergic projection in 3_D. Our findings show that serotonergic fibers projecting to olfactory bulb require a special timing to enter the target. The expression pattern of ephrinA5 suggests that ephrinA5 can be one of the factors that modulate this timing. Overall, our results show for the first time the implication a guidance molecule for the region-specific and time-specific targeting of serotonin raphe neurons and has implications for the anatomo-functional parsing of raphe cell groups.

GABAergic modulation of serotonergic neurons in the dorsal raphe nucleus

Article

Full-text available

Sep 2018

The dorsal raphe nucleus (DRN), located in the brainstem, is involved in several functions such as sleep, temperature regulation, stress responses, and anxiety behaviors. This nucleus contains the largest population of serotonin expressing neurons in the brain. Serotonergic DRN neurons receive tonic γ-aminobutyric acid (GABA)inhibitory inputs from several brain areas, as well as from interneurons within the same nucleus. Serotonergic and GABAergic neurons in the DRN can be distinguished by their size, location, pharmacological responses, and electrophysiological properties. GABAergic neurons regulate the excitability of DRN serotonergic neurons and the serotonin release in different brain areas. Also, it has been shown that GABAergic neurons can synchronize the activity of serotonergic neurons across functions such as sleep or alertness. Moreover, dysregulation of GABA signaling in the DRN has been linked to psychiatric disorders such as anxiety and depression. This review focuses on GABAergic transmission in the DRN. The interaction between GABAergic and serotonergic neurons is discussed considering some physiological implications. Also, the main electrophysiological and morphological characteristics of serotonergic and GABAergic neurons are described.

Circuitry Underlying Regulation of the Serotonergic System by Swim Stress

Article

Feb 2003

The dorsal raphe nucleus (DR)-serotonin (5-HT) system has been implicated in depression and is dramatically affected by swim stress, an animal model with predictive value for antidepressants. Accumulating evidence implicates the stress-related neuropeptide corticotropin-releasing factor (CRF) in the effect of swim stress on this system. This study investigated neural circuits within the DR that are activated by swim stress as revealed by neuronal expression of the immediate early gene, c-fos. Swim stress increased c-fos expression in the dorsolateral subregion of the DR. The majority of c-fos-expressing neurons were doubly labeled for GABA (85 +/- 5%), whereas relatively few were immunolabeled for 5-HT (4 +/- 1%), glutamate (0.5 +/- 0.3%) or calbindin (1.5 +/- 0.3%). Dual immunohistochemical labeling revealed that c-fos-expressing neurons in the dorsolateral DR were enveloped by dense clusters of CRF-immunoreactive fibers and also contained immunolabeling for CRF receptor, suggesting that c-fos-expressing neurons in the DR were specifically targeted by CRF. Consistent with this, the CRF receptor 1 antagonist, antalarmin, prevented swim-stress-elicited c-fos expression in the dorsolateral DR. Together with previous findings that both swim stress and CRF decrease 5-HT release in certain forebrain regions, these results suggest that swim stress engages CRF inputs to GABA neurons in the dorsolateral DR that function to inhibit 5-HT neurons and 5-HT release in the forebrain. This circuitry may underlie some of the acute behavioral responses to swim stress as well as the neuronal plasticity involved in long-term behavioral changes produced by this stress.

Serotonin neurons in the dorsal raphe mediate the anticataplectic action of orexin neurons by reducing amygdala activity

Article

Apr 2017

Significance Although the neurodegeneration of orexin (hypocretin)-producing neurons clearly causes the sleep disorder narcolepsy, the precise neural mechanisms by which orexin neurons prevent narcolepsy remain unclear. We previously demonstrated that orexin neurons inhibit cataplexy-like episodes—cataplexy is a cardinal symptom of narcolepsy, characterized by a sudden weakening of muscle tone—via serotonin neurons in the dorsal raphe nucleus (DRN). We thus used optogenetic and chemogenetic approaches to demonstrate that DRN serotonin neurons suppress cataplexy-like episodes by reducing the activity of the amygdala that plays an important role in emotional processing, as consistent with the fact that strong emotions often trigger cataplexy. We therefore propose that the orexin neuron–DRN serotonin neuron–amygdala pathway is a critical circuit for preventing cataplexy.

Constitutive and Acquired Serotonin Deficiency Alters Memory and Hippocampal Synaptic Plasticity

Article

Full-text available

Jul 2016
NEUROPSYCHOPHARMACOL

Serotonin (5-HT) deficiency occurs in a number of brain disorders that affect cognitive function. However, a direct causal relationship between 5-HT hypo-transmission and memory, and underlying mechanisms, has not been established. We used mice with a constitutive depletion of 5-HT brain levels, (Pet1KO mice) to analyze the contribution of 5-HT to different forms of learning and memory. Pet1KO mice exhibited a striking deficit in novel object recognition memory, a hippocampal-dependent task. No alterations were found in tasks for social recognition, procedural learning or fear memory. Viral delivery of Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) was used to selectively silence the activity of 5-HT neurons in the raphe. Inhibition of 5-HT neurons in the median raphe, but not the dorsal raphe, was sufficient to impair object recognition in adult mice. In vivo electrophysiology in behaving mice showed that long-term potentiation in the hippocampus of 5-HT deficient mice was altered, and administration of the 5-HT1A agonist 8-OHDPAT rescued the memory deficits. Our data suggest that hyposerotonergia selectively affects declarative hippocampal-dependent memory. Serotonergic projections from the median raphe are necessary to regulate object memory and hippocampal synaptic plasticity processes, through an inhibitory control mediated by 5-HT1A receptors.Neuropsychopharmacology accepted article preview online, 27 July 2016. doi:10.1038/npp.2016.134.

Voluntary activation of muscle in humans: does serotonergic neuromodulation matter?

Article

Full-text available

Aug 2022
J PHYSIOL-LONDON

Ionotropic inputs to motoneurones have the capacity to depolarise and hyperpolarise the motoneurone, whereas neuromodulatory inputs control the state of excitability of the motoneurone. Intracellular recordings of motoneurones from in vitro and in situ animal preparations have provided extraordinary insight into the mechanisms that underpin how neuromodulators regulate neuronal excitability. However, far fewer studies have attempted to translate the findings from cellular and molecular studies into a human model. In this review, we focus on the role that serotonin (5‐HT) plays in muscle activation in humans. 5‐HT is a potent regulator of neuronal firing rates, which can influence the force that can be generated by muscles during voluntary contractions. We firstly outline structural and functional characteristics of the serotonergic system, and then describe how motoneurone discharge can be facilitated and suppressed depending on the 5‐HT receptor subtype that is activated. We then provide a narrative on how 5‐HT effects can influence voluntary activation during muscle contractions in humans, and detail how 5‐HT may be a mediator of exercise‐induced fatigue that arises from the central nervous system. image

Functional connectome of arousal and motor brainstem nuclei in living humans by 7 Tesla resting-state fMRI

Article

Jan 2022
NEUROIMAGE

Brainstem nuclei play a pivotal role in many functions, such as arousal and motor control. Nevertheless, the connectivity of arousal and motor brainstem nuclei is understudied in living humans due to the limited sensitivity and spatial resolution of conventional imaging, and to the lack of atlases of these deep tiny regions of the brain. For a holistic comprehension of sleep, arousal and associated motor processes, we investigated in 20 healthy subjects the resting-state functional connectivity of 18 arousal and motor brainstem nuclei in living humans. To do so, we used high spatial-resolution 7 Tesla resting-state fMRI, as well as a recently developed in-vivo probabilistic atlas of these nuclei in stereotactic space. Further, we verified the translatability of our brainstem connectome approach to conventional (e.g. 3 Tesla) fMRI. Arousal brainstem nuclei displayed high interconnectivity, as well as connectivity to the thalamus, hypothalamus, basal forebrain and frontal cortex, in line with animal studies and as expected for arousal regions. Motor brainstem nuclei showed expected connectivity to the cerebellum, basal ganglia and motor cortex, as well as high interconnectivity. Comparison of 3 Tesla to 7 Tesla connectivity results indicated good translatability of our brainstem connectome approach to conventional fMRI, especially for cortical and subcortical (non-brainstem) targets and to a lesser extent for brainstem targets. The functional connectome of 18 arousal and motor brainstem nuclei with the rest of the brain might provide a better understanding of arousal, sleep and accompanying motor function in living humans in health and disease.

Effect of water restriction stress on social behavior in mice: Impact on serotonergic system plasticity

Article

Feb 2021

Die Auswirkungen von Tph2-Defizienz und negativen frühen Umwelterfahrungen auf Angstverhalten in weiblichen Mäusen

Thesis

Full-text available

Jan 2021

Charlotte Sophie Auth

Angsterkrankungen gehören zu den am weitesten verbreiteten psychischen Erkrankungen und stellen eine beträchtliche soziale und wirtschaftliche Herausforderung für unsere Gesellschaft dar. Aversive frühe Erfahrungen sind ein bekannter Risikofaktor für die Entwicklung verschiedener psychischer Erkrankungen, insbesondere Angststörungen. Während der frühen Entwicklung findet die Programmierung der Hypothalamus-Hypophysen-Nebennierenrinden- (HHN)-Achse, die die Ausschüttung des Stresshormons Cortisol in Menschen bzw. Corticosteron in Mäusen steuert, statt. Wenn Individuen in dieser kritischen Phase Stress ausgesetzt sind, wird die regelrechte Ausbildung der HHN-Achse gestört, was zu dysregulierten Verhaltensantworten auf Stressreize im späteren Leben führen kann. Das Serotonin (5-HT)-System als eines der ausgedehntesten Neurotransmittersysteme ist an der Vermittlung der Effekte von früher Stressexposition auf angstähnliche Verhaltensweisen beteiligt. Das Ziel dieser Studie ist es, die Interaktion zwischen genetischer Prädisposition und negativen Einflüssen in frühen Entwicklungsstadien auf die Ausbildung von Angstverhalten im Erwachsenenalter näher zu beleuchten. In dieser Studie wurden Tryptophanhydroxylase 2 (Tph2)-defiziente weibliche Mäuse als Modell für ein lebenslanges konstitutives 5-HT Synthesedefizit im zentralen Nervensystem verwendet. Nachkommen dieser Mauslinie wurden im frühen Lebensalter Maternaler Separation (MS), d.h. einem mütterlichen Trennungsparadigma, unterzogen und im Erwachsenenalter im „Open field“ (OF) oder in der „Dark-light box“ (DLB) getestet. Im Anschluss an die Verhaltensexperimente wurde die neuronale Aktivierung immunhistochemisch durch Darstellung des frühzeitig auftretenden Genprodukts c-Fos bestimmt. In der DLB zeigten homozygot Tph2-defiziente Mäuse eine verringerte motorische Aktivität im hellen Kompartiment, und dieser Effekt konnte durch MS normalisiert werden. Zusätzlich verstärkte MS bei diesem Genotyp das Auftreten von fluchtartigen Sprüngen. Im OF hat MS fluchtartige Verhaltensweisen in homo- und heterozygoten Tph2-defizienten Mäusen befördert. Beide Verhaltenstests führten zu spezifischen neuronalen Aktivierungsmustern, die mithilfe von c-Fos- Immunhistochemie ausgewertet wurden. Die Durchführung des DLB-Tests führte in Abhängigkeit vom Vorhandensein von Tph2 zur Aktivierung des paraventrikulären Kerns des Hypothalamus (PVN) und der basolateralen Amygdala (BL), wohingegen die Exposition gegenüber dem OF-Test zu einer Aktivierung der lateralen Amygdala (La) in Tieren, die einem mütterlichen Trennungsparadigma unterzogen wurden, sowie einer Aktivierung des ventrolateralen (VLPAG) und dorsolateralen (DLPAG) periaquäduktalen Höhlengraus in Abhängigkeit von Tph2 und MS führte. Zusammenfassend weisen die Ergebnisse dieser Studie darauf hin, dass MS aktive Verhaltensantworten auf aversive Reize in Abhängigkeit vom Vorhandensein von 5-HT im Gehirn fördert. Diese Effekte könnten durch die spezifische Aktivierung von mit Angstverhalten in Zusammenhang stehenden Gehirnregionen während der Verhaltensexperimente vermittelt werden.

Deciphering an AgRP-serotoninergic neural circuit in distinct control of energy metabolism from feeding

Article

Full-text available

Jun 2021

Contrasting to the established role of the hypothalamic agouti-related protein (AgRP) neurons in feeding regulation, the neural circuit and signaling mechanisms by which they control energy expenditure remains unclear. Here, we report that energy expenditure is regulated by a subgroup of AgRP neurons that send non-collateral projections to neurons within the dorsal lateral part of dorsal raphe nucleus (dlDRN) expressing the melanocortin 4 receptor (MC4R), which in turn innervate nearby serotonergic (5-HT) neurons. Genetic manipulations reveal a bi-directional control of energy expenditure by this circuit without affecting food intake. Fiber photometry and electrophysiological results indicate that the thermo-sensing MC4RdlDRN neurons integrate pre-synaptic AgRP signaling, thereby modulating the post-synaptic serotonergic pathway. Specifically, the MC4RdlDRN signaling elicits profound, bi-directional, regulation of body weight mainly through sympathetic outflow that reprograms mitochondrial bioenergetics within brown and beige fat while feeding remains intact. Together, we suggest that this AgRP neural circuit plays a unique role in persistent control of energy expenditure and body weight, hinting next-generation therapeutic approaches for obesity and metabolic disorders. Neuronal signaling has an important role in the regulation of energy expenditure and body weight, however, the underlying mechanisms are incompletely understood. Here, the authors report a AgRP-MC4R-serotonin expressing neuronal circuit that regulate energy expenditure without affecting feeding.

Migraine Is More Than Just Headache: Is the Link to Chronic Fatigue and Mood Disorders Simply Due to Shared Biological Systems?

Article

Full-text available

Jun 2021
FRONT HUM NEUROSCI

Migraine is a symptomatically heterogeneous condition, of which headache is just one manifestation. Migraine is a disorder of altered sensory thresholding, with hypersensitivity among sufferers to sensory input. Advances in functional neuroimaging have highlighted that several brain areas are involved even prior to pain onset. Clinically, patients can experience symptoms hours to days prior to migraine pain, which can warn of impending headache. These symptoms can include mood and cognitive change, fatigue, and neck discomfort. Some epidemiological studies have suggested that migraine is associated in a bidirectional fashion with other disorders, such as mood disorders and chronic fatigue, as well as with other pain conditions such as fibromyalgia. This review will focus on the literature surrounding alterations in fatigue, mood, and cognition in particular, in association with migraine, and the suggested links to disorders such as chronic fatigue syndrome and depression. We hypothesize that migraine should be considered a neural disorder of brain function, in which alterations in aminergic networks integrating the limbic system with the sensory and homeostatic systems occur early and persist after headache resolution and perhaps interictally. The associations with some of these other disorders may allude to the inherent sensory sensitivity of the migraine brain and shared neurobiology and neurotransmitter systems rather than true co-morbidity.

Effects of water restriction on social behavior and 5-HT neurons density in the dorsal and median raphe nuclei in mice

Article

Nov 2020
BEHAV BRAIN RES

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.

An investigation of changes in NMDA-receptor evoked monoamine efflux following administration of antidepressant drugs: a microdialysis study in vivo

Thesis

Jan 2003

Faddy Sadideen

It is widely accepted that the symptoms of depression are due, in part, to abnormal monoaminergic tone in the brain, primarily serotonin, noradrenaline and to a lesser extent dopamine. This constitutes the monoamine theory of depression. Antidepressants (ADs) work by increasing the extracellular concentration of monoamines at the synapse. Though, their mechanism is not fully understood, it has been suggested that chronic AD treatments can affect NMDA receptor function in the brain. Using in vivo microdialysis in freely moving rats, the effects of acute, 7-day subchronic and chronic doses of the ADs paroxetine and clomipramine treatment on the NMDA- evoked efflux of extracellular DA, 5-HT and their metabolites, DOPAC and 5-HIAA respectively in the frontal cortex were investigated. The duration of these effects after 48 hours and 14 days of drug cessation, and the effect of the co-administration of NMDA antagonists with paroxetine on monoamine levels and their metabolites was also investigated. Acute injection of paroxetine (10 and 20 mg/kg i.p.) did not affect dialysate DA or 5-HT content in the frontal cortex. Clomipramine at 10 and 20 mg/kg caused a decrease in extracellular DA without exerting any influence on dialysate 5-HT levels. Local infusion of 100μM NMDA into the frontal cortex decreased both extracellular DA and 5-HT levels in this region. 21 day treatment of rats with paroxetine and clomipramine increased 5-HT levels to 150% and 147% above basal levels respectively. The same treatment increased DA levels to 200% and 186% above basal levels. When NMDA infusion was preceded by a single injection of paroxetine/clomipramine no marked differences between NMDA and NMDA+paroxetine/clomipramine treated groups were observed. Subchronic (7-days) and chronic (21-days) treatment with paroxetine/clomipramine were able to abolish the NMDA-evoked decrease in dialysate DA and 5-HT levels. This effect lasted for a period of 48 hours but was abolished following a 14-day 'drug holiday'. This suggests that adaptive functional changes occur in NMDA receptor function during treatment with AD drugs. These results suggest that the NMDA receptor is subject to adaptive changes following chronic AD treatment. Interestingly, the co-administration of acute paroxetine with NMDA antagonists (amantadine, budipine, CGP 40116 and ifenprodil) causes an increase in extracellular 5-HT which may prove to have clinical implications.

Serotonergic innervation of the auditory midbrain: dorsal raphe subregions differentially project to the auditory midbrain in male and female mice

Article

Full-text available

Jul 2020
BRAIN STRUCT FUNCT

In the auditory inferior colliculus (IC), serotonin reflects features of context including the valence of social interactions, stressful events, and prior social experience. However, within the dorsal raphe nucleus (DRN; B6 + B7), the source of serotonergic projections to the IC has not been resolved at the level of DRN subregions. Additionally, few studies have investigated which DRN subregions are engaged during naturalistic, sensory-driven social behaviors. We employ traditional, retrograde tract-tracing approaches to comprehensively map the topographic extent of DRN-IC projection neurons in male and female mice. We combine this approach with immediate early gene (cFos) mapping in order to describe the functional properties of DRN subregions during contexts in which serotonin fluctuates within the IC. These approaches provide novel evidence that the dorsal (DRd) and lateral (DRl) B7 subregions are primarily responsible for serotonergic innervation of the IC; further, we show that this projection is larger in male than in female mice. Additionally, DRd and the ventral B7 (DRv) contained more transcriptionally active serotonergic neurons irrespective of behavioral context. Male mice had more active serotonergic neurons in DRd and DRv than females following sociosexual encounters. However, serotonergic activity was correlated with the expression of female but not male social behaviors. The topographic organization of the DRN-IC projection provides the anatomical framework to test a mechanism underlying context-dependent auditory processing. We further highlight the importance of including sex as a biological variable when describing the functional topography of DRN.

Hypoxia-ischemia in the immature rodent brain impairs serotonergic neuronal function in certain dorsal raphé nuclei

Article

Full-text available

Mar 2020

Neonatal hypoxia-ischemia (HI) results in losses of serotonergic neurons in specific dorsal raphé nuclei. However, not all serotonergic raphé neurons are lost and it is therefore important to assess the function of remaining neurons in order to understand their potential to contribute to neurological disorders in the HI-affected neonate. The main objective of this study was to determine how serotonergic neurons, remaining in the dorsal raphé nuclei after neonatal HI, respond to an external stimulus (restraint stress). On postnatal day 3 (P3), male rat pups were randomly allocated to one of the following groups: (i) control + no restraint (n = 5), (ii) control + restraint (n = 6), (iii) P3 HI + no restraint (n = 5) or (iv) P3 HI + restraint (n = 7). In the two HI groups, rat pups underwent surgery to ligate the common carotid artery and were then exposed to 6% O2 for 30 minutes. Six weeks after P3 HI, on P45, rats were subjected to restraint stress for 30 minutes. Using dual immunolabeling for Fos protein, a marker for neuronal activity, and serotonin (5-hydroxytrypamine; 5-HT), numbers of Fos-positive 5-HT neurons were determined in five dorsal raphé nuclei. We found that restraint stress alone increased numbers of Fos-positive 5-HT neurons in all five dorsal raphé nuclei compared to control animals. However, following P3 HI, the number of stress-induced Fos-positive 5-HT neurons was decreased significantly in the dorsal raphé ventrolateral, interfascicular and ventral nuclei compared with control animals exposed to restraint stress. In contrast, numbers of stress-induced Fos-positive 5-HT neurons in the dorsal raphé dorsal and caudal nuclei were not affected by P3 HI. These data indicate that not only are dorsal raphé serotonergic neurons lost after neonatal HI, but also remaining dorsal raphé serotonergic neurons have reduced differential functional viability in response to an external stimulus. Procedures were approved by the University of Queensland Animal Ethics Committee (UQCCR958/08/NHMRC) on February 27, 2009.

Activity-dependent heterogeneous populations of nitric oxide synthase neurons in the rat dorsal raphe nucleus

Research

Full-text available

Apr 2006

Lurasidone Sub-Chronically Activates Serotonergic Transmission via Desensitization of 5-HT1A and 5-HT7 Receptors in Dorsal Raphe Nucleus

Article

Full-text available

Oct 2019

Lurasidone is an atypical mood-stabilizing antipsychotic agent with unique receptor-binding profile, including 5-HT7 receptor (5-HT7R) antagonism. Effects of 5-HT7R antagonism on transmitter systems of schizophrenia and mood disorders, however, have not been well clarified. Thus, this study examined the mechanisms underlying the clinical effects of lurasidone by measuring mesocortical serotonergic transmission. Following systemic and local administrations of lurasidone, MK801 and 5-HT receptor modulators, we determined releases of 5-HT in dorsal raphe nucleus (DRN), mediodorsal thalamic nucleus (MDTN) and medial prefrontal cortex (mPFC) and γ-aminobutyric acid (GABA) in DRN using multiprobe microdialysis with ultra-high-performance liquid chromatography (UHPLC). Serotonergic and GABAergic neurons in the DRN are predominantly regulated by inhibitory 5-HT1A receptor (5-HT1AR) and excitatory 5-HT7R, respectively. Lurasidone acutely generates GABAergic disinhibition by 5-HT7R antagonism, but concomitant its 5-HT1AR agonism prevents serotonergic hyperactivation induced by 5-HT7R inhibition. During treatments with 5-HT1AR antagonist in DRN, lurasidone dose-dependently increased 5-HT release in the DRN, MDTN and mPFC. Contrary, lurasidone chronically enhanced serotonergic transmission and GABAergic disinhibition in the DRN by desensitizing both 5-HT1AR and 5-HT7R. These effects of lurasidone acutely prevented MK801-evoked 5-HT release by GABAergic disinhibition via N-methyl-D-aspartate (NMDA)/glutamate receptor (NMDA-R)-mediated inhibition of 5-HT1AR function, but enhanced MK801-induced 5-HT release by desensitizing 5-HT1AR and 5-HT7R. These results indicate that acutely lurasidone fails to affect 5-HT release, but chronically enhances serotonergic transmission by desensitizing both 5-HT1AR and 5-HT7R. These unique properties of lurasidone ameliorate the dysfunctions of NMDA-R and augment antidepressive effects.

Current understanding of fear learning and memory in humans and animal models and the value of a linguistic approach for analyzing fear learning and memory in humans

Article

Full-text available

Apr 2019

Fear is an emotion that serves as a driving factor in how organisms move through the world. In this review, we discuss the current understandings of the subjective experience of fear and the related biological processes involved in fear learning and memory. We first provide an overview of fear learning and memory in humans and animal models, encompassing the neurocircuitry and molecular mechanisms, the influence of genetic and environmental factors, and how fear learning paradigms have contributed to treatments for fear-related disorders, such as posttraumatic stress disorder. Current treatments as well as novel strategies, such as targeting the perisynaptic environment and use of virtual reality, are addressed. We review research on the subjective experience of fear and the role of autobiographical memory in fear-related disorders. We also discuss the gaps in our understanding of fear learning and memory, and the degree of consensus in the field. Lastly, the development of linguistic tools for assessments and treatment of fear learning and memory disorders is discussed.

Involvement of Serotonergic and Relaxin-3 Neuropeptide Systems in the Expression of Anxiety-like Behavior

Article

Aug 2018
NEUROSCIENCE

Anxiety-related defensive behavior is controlled by a distributed network of brain regions and interconnected neural circuits. The dorsal raphe nucleus (DR), which contains the majority of forebrain-projecting serotonergic neurons, is a key brain region involved in fear states and anxiety-related behavior via modulation of this broad neural network. Evidence suggests that relaxin-3 neurons in the nucleus incertus (NI) may also interact with this network, however, the potential role of the NI in the control of anxiety-related defensive behavior requires further investigation. In this study, we examined the response of an anxiety-related neuronal network, including serotonergic neurons in the DR and relaxin-3-containing neurons in the NI, to administration of an anxiogenic drug and exposure to an aversive environment. We administered an anxiogenic dose of the adenosine receptor antagonist, caffeine (50 mg/kg, i.p.), or vehicle, to adult male Wistar rats and 30 min later exposed them to either an elevated plus-maze (EPM) or a home-cage environment. Administration of caffeine and exposure to the EPM activated a broad network of brain regions involved in control of anxiety-like behaviors, including serotonergic neurons in the DR, as measured using c-Fos immunohistochemistry. However, only exposure to the EPM activated relaxin-3-containing neurons in the NI, and activation of these neurons was not correlated with changes in anxiety-like behavior. These data suggest activation of the NI relaxin-3 system is associated with expression of behavior in tests of anxiety, but may not be directly involved in the approach-avoidance conflict inherent in anxiety-related defensive behavior in rodents.

5-HT2C receptor blockade reverses SSRI-associated basal ganglia dysfunction and potentiates therapeutic efficacy

Article

Full-text available

Dec 2020

Serotonin (5-HT) selective reuptake inhibitors (SSRIs) are widely used in the treatment of depression and anxiety disorders, but responsiveness is uncertain and side effects often lead to discontinuation. Side effect profiles suggest that SSRIs reduce dopaminergic (DAergic) activity, but specific mechanistic insight is missing. Here we show in mice that SSRIs impair motor function by acting on 5-HT2C receptors in the substantia nigra pars reticulata (SNr), which in turn inhibits nigra pars compacta (SNc) DAergic neurons. SSRI-induced motor deficits can be reversed by systemic or SNr-localized 5-HT2C receptor antagonism. SSRIs induce SNr hyperactivity and SNc hypoactivity that can also be reversed by systemic 5-HT2C receptor antagonism. Optogenetic inhibition of SNc DAergic neurons mimics the motor deficits due to chronic SSRI treatment, whereas local SNr 5-HT2C receptor antagonism or optogenetic activation of SNc DAergic neurons reverse SSRI-induced motor deficits. Lastly, we find that 5-HT2C receptor antagonism potentiates the antidepressant and anxiolytic effects of SSRIs. Together our findings demonstrate opposing roles for 5-HT2C receptors in the effects of SSRIs on motor function and affective behavior, highlighting the potential benefits of 5-HT2C receptor antagonists for both reduction of motor side effects of SSRIs and augmentation of therapeutic antidepressant and anxiolytic effects.

Circuit mechanisms and computational models of REM sleep

Article

Aug 2018
NEUROSCI RES

Rapid eye movement (REM) sleep or paradoxical sleep is an elusive behavioral state. Since its discovery in the 1950s, our knowledge of the neuroanatomy, neurotransmitters and neuropeptides underlying REM sleep regulation has continually evolved in parallel with the development of novel technologies. Although the pons was initially discovered to be responsible for REM sleep, it has since been revealed that many components in the hypothalamus, midbrain, pons, and medulla also contribute to REM sleep. In this review, we first provide an up-to-date overview of REM sleep-regulating circuits in the brainstem and hypothalamus by summarizing experimental evidence from neuroanatomical, neurophysiological and gain- and loss-of-function studies. Second, because quantitative approaches are essential for understanding the complexity of REM sleep-regulating circuits and because mathematical models have provided valuable insights into the dynamics underlying REM sleep genesis and maintenance, we summarize computational studies of the sleep-wake cycle, with an emphasis on REM sleep regulation. Finally, we discuss outstanding issues for future studies.

Brain structure and function related to headache: Brainstem structure and function in headache

Article

Jul 2018

Objective: To review and discuss the literature relevant to the role of brainstem structure and function in headache. Background: Primary headache disorders, such as migraine and cluster headache, are considered disorders of the brain. As well as head-related pain, these headache disorders are also associated with other neurological symptoms, such as those related to sensory, homeostatic, autonomic, cognitive and affective processing that can all occur before, during or even after headache has ceased. Many imaging studies demonstrate activation in brainstem areas that appear specifically associated with headache disorders, especially migraine, which may be related to the mechanisms of many of these symptoms. This is further supported by preclinical studies, which demonstrate that modulation of specific brainstem nuclei alters sensory processing relevant to these symptoms, including headache, cranial autonomic responses and homeostatic mechanisms. Review focus: This review will specifically focus on the role of brainstem structures relevant to primary headaches, including medullary, pontine, and midbrain, and describe their functional role and how they relate to mechanisms of primary headaches, especially migraine.

Social approach, anxiety, and altered tryptophan hydroxylase 2 activity in juvenile BALB/c and C57BL/6J mice

Article

Jun 2018
BEHAV BRAIN RES

Autism spectrum disorder (ASD) is a heterogeneous and highly heritable condition with multiple aetiologies. Although the biological mechanisms underlying ASD are not fully understood, evidence suggests that dysregulation of serotonergic systems play an important role in ASD psychopathology. Preclinical models using mice with altered serotonergic neurotransmission may provide insight into the role of serotonin in behaviours relevant to clinical features of ASD. For example, BALB/c mice carry a loss-of-function single nucleotide polymorphism (SNP; C1473 G) in tryptophan hydroxylase 2 (Tph2), which encodes the brain-specific isoform of the rate-limiting enzyme for serotonin synthesis, and these mice frequently have been used to model symptoms of ASD. In this study, juvenile male BALB/c (G/G; loss-of-function variant) and C57BL/6 J (C/C; wild type variant) mice, were exposed to the three-chamber sociability test, and one week later to the elevated plus-maze (EPM). Tryptophan hydroxylase 2 (TPH2) activity was measured following injection of the aromatic amino acid decarboxylase (AADC)-inhibitor, NSD-1015, and subsequent HPLC detection of 5-hydroxytryptophan (5-HTP) within subregions of the dorsal raphe nucleus (DR) and median raphe nucleus (MnR). The BALB/c mice showed reduced social behaviour and increased anxious behaviour, as well as decreased 5-HTP accumulation in the rostral and mid-rostrocaudal DR. In the full cohort of mice, TPH2 activity in the mid-rostrocaudal DR was correlated with anxious behaviour in the EPM, however these correlations were not statistically significant within each strain, suggesting that TPH2 activity was not directly associated with either anxiety or sociability. Further research is therefore required to more fully understand how serotonergic systems are involved in mouse behaviours that resemble some of the clinical features of ASD.

The Role of Corticotropin-Releasing Hormone in the Dorsal Raphe Nucleus in Mediating the Behavioral Consequences of Uncontrollable Stress

Article

Feb 2002

Serotonin selectively attenuates glutamate-evoked activation of noradrenergic locus coeruleus neurons

Article

Full-text available

Mar 1991

The effect of 5-HT on activity of noradrenergic locus coeruleus (LC) neurons was studied using microiontophoretic and micropressure drug application in anesthetized rats. 5-HT had no consistent effect on LC spontaneous discharge, eliciting a modest decrease overall. However, 5- HT reliably attenuated responses of LC neurons to excitatory amino acids (EAAs), one of the major classes of transmitters in afferents to these neurons. This effect was specific for EAA responses because it occurred for glutamate and kainate but not for ACh. In contrast, iontophoretic norepinephrine (NE) selectively attenuated spontaneous activity but not responses evoked by either glutamate or ACh. The responsiveness of LC neurons to EAAs as quantified by a response- contrast measure (evoked excitation/basal activity) was markedly reduced by 5-HT, but was increased by NE. For ACh, such responsiveness of LC cells was not changed by 5-HT, but was increased by NE. The effects of 5-HT were prevented and reversed by iontophoretically applied antagonists of 5-HT receptors, methysergide and methiothepin. Thus, 5-HT appears to selectively interact with EAA responses of LC neurons, acting as a filter to attenuate LC activity linked to its major EAA inputs while allowing other channels afferent to the LC (e.g., those utilizing ACh) to be expressed.

GluA2-Lacking AMPA Receptors and Nitric Oxide Signaling Gate Spike-Timing Dependent Potentiation of Glutamate Synapses in the Dorsal Raphe Nucleus

Article

Full-text available

May 2017

The dorsal raphe nucleus (DRn) receives glutamatergic inputs from numerous brain areas that control the function of DRn serotonin (5-HT) neurons. By integrating these synaptic inputs, 5-HT neurons modulate a plethora of behaviors and physiological functions. However, it remains unknown whether the excitatory inputs onto DRn 5-HT neurons can undergo activity-dependent change of strength, as well as the mechanisms that control their plasticity. Here, we describe a novel form of spike-timing–dependent long-term potentiation (tLTP) of glutamate synapses onto rat DRn 5-HT neurons. This form of synaptic plasticity is initiated by an increase in postsynaptic intracellular calcium but is maintained by a persistent increase in the probability of glutamate release. The tLTP of glutamate synapses onto DRn 5-HT is independent of NMDA receptors but requires the activation of calcium-permeable AMPA receptors and voltage-dependent calcium channels. The presynaptic expression of the tLTP is mediated by the retrograde messenger nitric oxide (NO) and activation of cGMP/PKG pathways. Collectively, these results indicate that glutamate synapses in the DRn undergo activity-dependent synaptic plasticity gated by NO signaling and unravel a previously unsuspected role of NO in controlling synaptic function and plasticity in the DRn.

Benefits of Hormone Therapy Estrogens Depend on Estrogen Type: 17β-Estradiol and Conjugated Equine Estrogens Have Differential Effects on Cognitive, Anxiety-Like, and Depressive-Like Behaviors and Increase Tryptophan Hydroxylase-2 mRNA Levels in Dorsal Raphe Nucleus Subregions

Article

Full-text available

Dec 2016

Decreased serotonin (5-HT) function is associated with numerous cognitive and affective disorders. Women are more vulnerable to these disorders and have a lower rate of 5-HT synthesis than men. Serotonergic neurons in the dorsal raphe nucleus (DRN) are a major source of 5-HT in the forebrain and play a critical role in regulation of stress-related disorders. In particular, polymorphisms of tryptophan hydroxylase-2 (TpH2, the brain-specific, rate-limiting enzyme for 5-HT biosynthesis) are implicated in cognitive and affective disorders. Administration of 17β-estradiol (E2), the most potent naturally circulating estrogen in women and rats, can have beneficial effects on cognitive, anxiety-like, and depressive-like behaviors. Moreover, E2 increases TpH2 mRNA in specific subregions of the DRN. Although conjugated equine estrogens (CEE) are a commonly prescribed estrogen component of hormone therapy in menopausal women, there is a marked gap in knowledge regarding how CEE affects these behaviors and the brain 5-HT system. Therefore, we compared the effects of CEE and E2 treatments on behavior and TpH2 mRNA. Female Sprague-Dawley rats were ovariectomized, administered either vehicle, CEE, or E2 and tested on a battery of cognitive, anxiety-like, and depressive-like behaviors. The brains of these animals were subsequently analyzed for TpH2 mRNA. Both CEE and E2 exerted beneficial behavioral effects, although efficacy depended on the distinct behavior and for cognition, on the task difficulty. Compared to CEE, E2 generally had more robust anxiolytic and antidepressant effects. E2 increased TpH2 mRNA in the caudal and mid DRN, corroborating previous findings. However, CEE increased TpH2 mRNA in the caudal and rostral, but not the mid, DRN, suggesting that distinct estrogens can have subregion-specific effects on TpH2 gene expression. We also found differential correlations between the level of TpH2 mRNA in specific DRN subregions and behavior, depending on the type of behavior. These distinct associations imply that cognition, anxiety-like, and depressive-like behaviors are modulated by unique serotonergic neurocircuitry, opening the possibility of novel avenues of targeted treatment for different types of cognitive and affective disorders.

Probing the neurophysiology of anxiety: Social stress alters the modulation of serotonin neurons

Article

Aug 2010

LaTasha K Crawford

Anxiety disorders are prevalent in human and veterinary medicine yet the underlying mechanism is poorly understood. Because serotonin (5-HT) neurons of the dorsal raphe (DR) are thought to play a prominent role, my goal was to understand the changes in DR 5-HT neurons that underlie anxiety and other stress-related disorders. Two DR subdivisions were studied in a series of experiments: the ventromedial DR (vmDR), a well characterized subfield with a high density of 5-HT neurons, and the lateral wing DR (lwDR), a largely uncharacterized subfield with a more sparse distribution of 5-HT neurons. Many stress paradigms activate 5-HT neurons of the lwDR more so than 5-HT neurons of the vmDR, suggesting a unique role for lwDR 5-HT cells in stress circuits. However, it is not known if lwDR 5-HT neurons possess physiological characteristics that contribute to their increased propensity to be activated by a stressor. I found that lwDR 5-HT neurons demonstrated increased intrinsic excitability, increased glutamatergic input, and similar GABAergic input when compared to vmDR 5-HT neurons. Using the chronic social defeat model of anxiety, the distinctions between lwDR and vmDR neurons were explored further. Social defeat induced anxious behavior and stress-associated pathological changes in the peripheral organs of intruder mice. For the first time, investigation into the neural mechanisms of social defeat has focused on 5-HT neuron physiology, revealing subregion-specific effects within the DR. Increased excitability was seen in the vmDR neurons of the most anxious mice. This was accompanied by a decrease in GABAergic input to vmDR 5-HT neurons potentially mediated by both presynaptic and postsynaptic changes. The lwDR 5-HT neurons demonstrated distinct stress-induced changes limited to the slower kinetics of postsynaptic GABAAR. The differential effect of social stress on inhibitory input to vmDR or lwDR neurons suggest that the 5-HT output in brain regions targeted by each subfield is differentially affected in anxiety disorders. Collectively these findings help fill the gap in our understanding of local DR circuitry, the heterogeneity of 5-HT neurons, and the distinct regulation of vmDR and lwDR neurons in the circuits that mediate stress and contribute to the pathophysiology of anxiety.

Tetratricopeptide repeat domain 9A modulates anxiety-like behavior in female mice

Article

Full-text available

Nov 2016

Tetratricopeptide repeat domain 9A (TTC9A) expression is abundantly expressed in the brain. Previous studies in TTC9A knockout (TTC9A−/−) mice have indicated that TTC9A negatively regulates the action of estrogen. In this study we investigated the role of TTC9A on anxiety-like behavior through its functional interaction with estrogen using the TTC9A−/− mice model. A battery of tests on anxiety related behaviors was conducted. Our results demonstrated that TTC9A−/− mice exhibited an increase in anxiety-like behaviors compared to the wild type TTC9A+/+ mice. This difference was abolished after ovariectomy, and administration of 17-β-estradiol benzoate (EB) restored this escalated anxiety-like behavior in TTC9A−/− mice. Since serotonin is well-known to be the key neuromodulator involved in anxiety behaviors, the mRNA levels of tryptophan hydroxylase (TPH) 1, TPH2 (both are involved in serotonin synthesis), and serotonin transporter (5-HTT) were measured in the ventromedial prefrontal cortex (vmPFC) and dorsal raphe nucleus (DRN). Interestingly, the heightened anxiety in TTC9A−/− mice under EB influence is consistent with a greater induction of TPH 2, and 5-HTT by EB in DRN that play key roles in emotion regulation. In conclusion, our data indicate that TTC9A modulates the anxiety-related behaviors through modulation of estrogen action on the serotonergic system in the DRN.

Serotonergic Neurons Are Targets for Leptin in the Monkey 1

Article

Jan 2001

Leptin is a secretory product of adipocytes that has been shown to affect food intake, metabolism, and reproduction. One site of leptin’s action is the central nervous system, where the leptin receptor (Ob-R) messenger ribonucleic acid (mRNA) and protein are expressed in discrete areas. In both the rat and monkey, Ob-R mRNA has been localized in the Raphe nuclei of the brainstem. Neurons in the Raphe nuclei are the primary source of serotonin in the brain. Serotonergic pathways influence both feeding and reproduction, and these cells are plausible direct targets for leptin’s action. We used double label in situ hybridization and computerized image analysis to determine whether serotonergic neurons in the brainstem of the female pigtailed macaque (Macaca nemestrina) express Ob-R mRNA. We observed that many cells in the Raphe nuclei express serotonin transporter mRNA, a marker of serotonergic cells, and Ob-R mRNA. Based on quantitative analysis, the highest number of cells that express both serotonin transporter and Ob-R mRNAs were found in the caudal dorsal Raphe and median Raphe nuclei; fewer double labeled cells were situated in the caudal linear nucleus and rostral median Raphe, whereas double labeled cells occurred infrequently in the rostral dorsal Raphe. These observations suggest that leptin may act on serotonergic cells to mediate some of its effects on ingestive behavior, metabolism, and reproduction. Leptin is a secretory product of adipocytes that has been shown to affect food intake, metabolism, and reproduction. One site of leptin’s action is the central nervous system, where the leptin receptor (Ob-R) messenger ribonucleic acid (mRNA) and protein are expressed in discrete areas. In both the rat and monkey, Ob-R mRNA has been localized in the Raphe nuclei of the brainstem. Neurons in the Raphe nuclei are the primary source of serotonin in the brain. Serotonergic pathways influence both feeding and reproduction, and these cells are plausible direct targets for leptin’s action. We used double label in situ hybridization and computerized image analysis to determine whether serotonergic neurons in the brainstem of the female pigtailed macaque (Macaca nemestrina) express Ob-R mRNA. We observed that many cells in the Raphe nuclei express serotonin transporter mRNA, a marker of serotonergic cells, and Ob-R mRNA. Based on quantitative analysis, the highest number of cells that express both serotonin transporter and Ob-R mRNAs were found in the caudal dorsal Raphe and median Raphe nuclei; fewer double labeled cells were situated in the caudal linear nucleus and rostral median Raphe, whereas double labeled cells occurred infrequently in the rostral dorsal Raphe. These observations suggest that leptin may act on serotonergic cells to mediate some of its effects on ingestive behavior, metabolism, and reproduction.

Monoaminergic Neuropathology in Alzheimer's disease

Article

Apr 2016

None of the proposed mechanisms of Alzheimer's disease (AD) fully explains the distribution patterns of the neuropathological changes at the cellular and regional levels, and their clinical correlates. One aspect of this problem lies in the complex genetic, epigenetic, and environmental landscape of AD: early-onset AD is often familial with autosomal dominant inheritance, while the vast majority of AD cases are late-onset, with the ε4 variant of the gene encoding apolipoprotein E (APOE) known to confer a 5-20 fold increased risk with partial penetrance. Mechanisms by which genetic variants and environmental factors influence the development of AD pathological changes, especially neurofibrillary degeneration, are not yet known. Here we review current knowledge of the involvement of the monoaminergic systems in AD. The changes in the serotonergic, noradrenergic, dopaminergic, histaminergic, and melatonergic systems in AD are briefly described. We also summarize the possibilities for monoamine-based treatment in AD. Besides neuropathologic AD criteria that include the noradrenergic locus coeruleus (LC), special emphasis is given to the serotonergic dorsal raphe nucleus (DRN). Both of these brainstem nuclei are among the first to be affected by tau protein abnormalities in the course of sporadic AD, causing behavioral and cognitive symptoms of variable severity. The possibility that most of the tangle-bearing neurons of the LC and DRN may release amyloid β as well as soluble monomeric or oligomeric tau protein trans-synaptically by their diffuse projections to the cerebral cortex emphasizes their selective vulnerability and warrants further investigations of the monoaminergic systems in AD.

Serotonin, Amygdala and Fear: Assembling the Puzzle

Article

Full-text available

Apr 2016

The fear circuitry orchestrates defense mechanisms in response to environmental threats. This circuitry is evolutionarily crucial for survival, but its dysregulation is thought to play a major role in the pathophysiology of psychiatric conditions in humans. The amygdala is a key player in the processing of fear. This brain area is prominently modulated by the neurotransmitter serotonin (5-hydroxytryptamine, 5-HT). The 5-HT input to the amygdala has drawn particular interest because genetic and pharmacological alterations of the 5-HT transporter (5-HTT) affect amygdala activation in response to emotional stimuli. Nonetheless, the impact of 5-HT on fear processing remains poorly understood.The aim of this review is to elucidate the physiological role of 5-HT in fear learning via its action on the neuronal circuits of the amygdala. Since 5-HT release increases in the basolateral amygdala (BLA) during both fear memory acquisition and expression, we examine whether and how 5-HT neurons encode aversive stimuli and aversive cues. Next, we describe pharmacological and genetic alterations of 5-HT neurotransmission that, in both rodents and humans, lead to altered fear learning. To explore the mechanisms through which 5-HT could modulate conditioned fear, we focus on the rodent BLA. We propose that a circuit-based approach taking into account the localization of specific 5-HT receptors on neurochemically-defined neurons in the BLA may be essential to decipher the role of 5-HT in emotional behavior. In keeping with a 5-HT control of fear learning, we review electrophysiological data suggesting that 5-HT regulates synaptic plasticity, spike synchrony and theta oscillations in the BLA via actions on different subcellular compartments of principal neurons and distinct GABAergic interneuron populations. Finally, we discuss how recently developed optogenetic tools combined with electrophysiological recordings and behavior could progress the knowledge of the mechanisms underlying 5-HT modulation of fear learning via action on amygdala circuits. Such advancement could pave the way for a deeper understanding of 5-HT in emotional behavior in both health and disease.

Prior stress and vasopressin promote corticotropin-releasing factor inhibition of serotonin release in the central nucleus of the amygdala

Article

Full-text available

Mar 2023

Corticotropin-releasing factor (CRF) is essential for coordinating endocrine and neural responses to stress, frequently facilitated by vasopressin (AVP). Previous work has linked CRF hypersecretion, binding site changes, and dysfunctional serotonergic transmission with anxiety and affective disorders, including clinical depression. Crucially, CRF can alter serotonergic activity. In the dorsal raphé nucleus and serotonin (5-HT) terminal regions, CRF effects can be stimulatory or inhibitory, depending on the dose, site, and receptor type activated. Prior stress alters CRF neurotransmission and CRF-mediated behaviors. Lateral, medial, and ventral subdivisions of the central nucleus of the amygdala (CeA) produce CRF and coordinate stress responsiveness. The purpose of these experiments was to determine the effect of intracerebroventricular (icv) administration of CRF and AVP on extracellular 5-HT as an index of 5-HT release in the CeA, using in vivo microdialysis in freely moving rats and high performance liquid chromatography (HPLC) analysis. We also examined the effect of prior stress (1 h restraint, 24 h prior) on CRF- and AVP-mediated release of 5-HT within the CeA. Our results show that icv CRF infusion in unstressed animals had no effect on 5-HT release in the CeA. Conversely, in rats with prior stress, CRF caused a profound dose-dependent decrease in 5-HT release within the CeA. This effect was long-lasting (240 min) and was mimicked by CRF plus AVP infusion without stress. Thus, prior stress and AVP functionally alter CRF-mediated neurotransmission and sensitize CRF-induced inhibition of 5-HT release, suggesting that this is a potential mechanism underlying stress-induced affective reactivity in humans.

Anatomical and neurochemical organization of the serotonergic system in the mammalian brain and in particular the involvement of the dorsal raphe nucleus in relation to neurological diseases

Chapter

Full-text available

Mar 2021
Progr Brain Res

The brainstem is a neglected brain area in neurodegenerative diseases, including Alzheimer's and Parkinson's disease, frontotemporal lobar degeneration and autonomic dysfunction. In Depression, several observations have been made in relation to changes in one particular the Dorsal Raphe Nucleus (DRN) which also points toward as key area in various age-related and neurodevelopmental diseases. The DRN is further thought to be related to stress regulated processes and cognitive events. It is involved in neurodegeneration, e.g., amyloid plaques, neurofibrillary tangles, and impaired synaptic transmission in Alzheimer's disease as shown in our autopsy findings. The DRN is a phylogenetically old brain area, with projections that reach out to a large number of regions and nuclei of the central nervous system, particularly in the forebrain. These ascending projections contain multiple neurotransmitters. One of the main reasons for the past and current interest in the DRN is its involvement in depression, and its main transmitter serotonin. The DRN also points toward the increased importance and focus of the brainstem as key area in various age-related and neurodevelopmental diseases. This review describes the morphology, ascending projections and the complex neurotransmitter nature of the DRN, stressing its role as a key research target into the neural bases of depression.

Midbrain circuits of novelty processing

Article

Dec 2020

Novelty triggers an increase in orienting behavior that is critical to evaluate the potential salience of unknown events. As novelty becomes familiar upon repeated encounters, this increase in response rapidly habituates as a form of behavioral adaptation underlying goal-directed behaviors. Many neurodevelopmental, psychiatric and neurodegenerative disorders are associated with abnormal responses to novelty and/or familiarity, although the neuronal circuits and cellular/molecular mechanisms underlying these natural behaviors in the healthy brain are largely unknown, as is the maladaptive processes that occur to induce impairment of novelty signaling in diseased brains. In rodents, the development of cutting-edge tools that allow for measurements of real time activity dynamics in selectively identified neuronal ensembles by gene expression signatures is beginning to provide advances in understanding the neural bases of the novelty response. Accumulating evidence indicate that midbrain circuits, the majority of which linked to dopamine transmission, promote exploratory assessments and guide approach/avoidance behaviors to different types of novelty via specific projection sites. The present review article focuses on midbrain circuit analysis relevant to novelty processing and habituation with familiarity.

Conséquences fonctionnelles, comportementales et adaptatives d'une mutation de la MAO (MonoAmine Oxydase) chez le poisson cavernicole aveugle Astyanax mexicanus.

Thesis

Jun 2020

Constance Pierre

Le neurotransmetteur sérotonine contrôle une grande variété de mécanismes physiologiques et comportementaux. Chez les humains, les mutations qui affectent la monoamine oxydase (MAO), l’enzyme qui dégrade la sérotonine, sont hautement délétères. Pourtant, le morphotype cavernicole aveugle de l’espèce Astyanax mexicanus (téléostéens) porte une mutation (P106L) induisant une perte de fonction partielle de cette enzyme, et semble prospérer dans son habitat souterrain naturel. Cette thèse décrit les effets de cette mutation, depuis l’échelle moléculaire à l’échelle des populations, afin de mieux cerner sa contribution à l’évolution des traits neuro-comportementaux du poisson cavernicole lors de son adaptation au milieu souterrain.Dans une première publication, nous avons établi 4 lignées de poissons, correspondant aux deux morphotypes de l’espèce Astyanax mexicanus (poisson cavernicole et poisson vivant en surface), chacun porteur ou non de la mutation P106L. La mutation P106L affecte le stress en augmentant l’amplitude de la réponse aigue au stress, tout en diminuant l’anxiété. Nous avons aussi étudié la distribution des allèles mutés dans les populations naturelles au Mexique et trouvé que l’allèle mutant est présent dans plusieurs grottes. L’évolution de l’allèle muté sous sélection ou dérive génétique est discutée.Dans une seconde publication, nous avons étudié les conséquences structurelles et biochimiques de la mutation P106L. La réduction de l’activité enzymatique de la MAO mutée est probablement due à une réduction de la flexibilité de l’une des boucles formant l’entrée au site actif, limitant l’accès des substrats. Des mesures d’HPLC ont montré des perturbations majeures de taux de sérotonine, dopamine et noradrénaline (et métabolites) dans le cerveau des poissons mutants. La mutation P106L mao est donc entièrement responsable du déséquilibre monoaminergique observé dans le cerveau des poissons cavernicoles. Enfin, l’effet de la mutation est partiellement compensé par une réduction de l’activité de synthèse de la sérotonine par la TPH. Nos résultats en révèlent plus sur les spécificités des systèmes monoaminergiques des poissons.Enfin, pour aggraver l’inhibition de la MAO, nous avons généré le premier poisson mutant knockout pour la MAO. Les mutants KO homozygotes sont chétifs et meurent durant les premières semaines de développement. Contrairement au système dopaminergique, le système sérotoninergique des poissons KO homozygotes est très fortement altéré : aucun neurone positif à la sérotonine n’est détecté dans l’hypothalamus.

Autoradiographic study of the distribution of 5-ht re-uptake sites in rat and human brain

Thesis

Jan 1995

Jacqueline Kay Miller

The selective serotonin re-uptake inhibitors (SSRI's) are a novel group of antidepressants which are rapidly becoming a popular choice of drug for the successful treatment of depression. The hypothesis of their mode of action, however, in conjunction with the cause of depression, remains subject to many inconsistencies. By using autoradiography to determine the distribution of the recognition sites of these drugs in the rat brain, it may be possible to detect discrete alterations in the serotonin re-uptake site which are not revealed in membrane binding studies. In the human brain, isolated areas of tissue have been used to provide some details of the sites of recognition of the SSRI's, however these studies have been limited by technical problems. Experimental protocols were established for the use of the SSRI 3H-paroxetine with autoradiography. The distribution of 3H-paroxetine binding sites in the rat brain revealed a heterogeneity of binding site densities, with the highest binding levels observed in the raphe nuclei in the brainstem. Direct injection of 5,7-dihydroxytryptamine (a selective serotonergic neurotoxin) into the dorsal raphe produced a reduction in 3H-paroxetine binding levels throughout the brain, thereby confirming the selectivity of 3H-paroxetine for the serotonin nerve terminal transporter site. A novel method of performing autoradiography in human whole brain sections was developed. By modifying a technique originally used for cutting whole body animal sections, it was possible to cryosection whole brain coronal slices (40μm) for use in autoradiography, whilst preserving the integrity of the brain tissue. The distribution of 3H-paroxetine binding sites within the human brain was determined and the symmetry of these binding sites established. A species difference in the localisation of the serotonin re-uptake site in rat and human brain was detected in a comparative study using 3H-paroxetine.

Activation of 5-HT1A receptors: Behavioural and neurochemical studies

Thesis

Jan 1997

Mark Thomas O'Connell

In rat brain the serotonin (5-hydroxytryptamine, 5-HT) receptor subtype, 5-HT1A receptor, is present at greatest density in the raphe nuclei on 5-HT cell bodies as a presynaptic autoreceptor, where its activation can decrease neuronal firing, reduce 5-HT synthesis, and decrease the release of 5-HT at terminal sites. The 5-HT1A receptor is also located postsynaptically where its activation produces behavioural responses such as components of the serotonergic syndrome. In this thesis the effects of 5-HT1A receptor activation by single and repeated administration of the selective agonist, 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH- DPAT) and by administration of the antagonist / partial agonist, 8-[2-[4-(2- methoxyphenyl)-1 -piperazinyl]ethyl]-8-azaspiro1[4,5]-decane-7,9-dione dihydrochloride (BMY 7378) were studied. Using a standardised experimental design, in which the effects of pretreatment with a 1 mg/kg s.c. dose of 8-OH-DPAT for 1,3 or 14 days on responses to a challenge dose of 0.5 mg/kg s.c. 8-OH-DPAT, the following results were obtained. Different behavioural responses to activation of 5-HT1A receptors by 8-OH-DPAT had strikingly different vulnerabilities to attenuation by 8-OH-DPAT pretreatment. Components of the 5-HT syndrome previously demonstrated to be mediated by postsynaptic 5-HT1A receptors were substantially attenuated after a single 8-OH-DPAT pretreatment. However another postsynaptic 5-HT1A receptor mediated response, tail-flick, and hyperphagia, a response to activation of presynaptic 5-HT1A receptors were both unaffected by 1,3 or 14 days pretreatment. The hypothermic effect of 8-OH-DPAT which, m the rat (as supported by evidence in this thesis), was shown to be a postsynaptically mediated 5-HT1A receptor response, was progressively attenuated over fourteen pretreatments. These results indicate that the process of desensitisation of responses to activation of the 5-HT1A receptor is complex and may be due to the presence of spare receptors (receptor reserve) or changes in secondary messenger interactions or by modulation of other downstream neurotransmitters. Evidence for a postsynaptic location for the 5-HT1A receptors mediating hypothermia was derived from results demonstrating that gross 5-HT depletion by the 5-HT synthesis inhibitor, p-chlorophenylalanine (pCPA) failed to attenuate the hypothermic effects of 8-OH-DPAT or BMY 7378. More convincing still was the finding that 8-OH-DPAT, a strong agonist at both presynaptic and postsynaptic 5-HT1A sites has comparable ED50 values for both hypothermia and hyperphagia, while BMY 7378, which is an agonist at presynaptic 5-HT1A receptors but mainly an antagonist at postsynaptic sites has an ED50 value for hypothermia which is two orders of magnitude greater than that for hyperphagia. In addition, infusion of BMY 7378 into the dorsal raphe was without clear hypothermic effect. The consequences of partial 5-HT depletion (circa 50%) following a single pCPA administration, on extracellular 5-HT in the frontal cortex, was investigated to assess the availability of 5-HT for release when the tissue content is lowered. Results indicate that the availability of 5-HT for release falls in proportion to the depletion of 5- HT stores. However, the resulting tissue 5-HT / 5-HIAA ratio would suggest that increased conservation of the partially depleted 5-HT stores may occur. In conclusion, the neurochemical and behavioural effects of activation of 5- HT1A receptors as indicated in this thesis are complex. Effects of this type as well as desensitisation of the 5-HT1A autoreceptor could play some part in antidepressant action.

Sensitive and Selective Measurement of Serotonin In Vivo Using Fast Cyclic Square-Wave Voltammetry

Article

Full-text available

Dec 2019

Although N-shaped fast scan cyclic voltammetry (N-FSCV) is well-established as an electroanalytical method to measure extracellular serotonin concentrations in vivo it is in need of improvement in both sensitivity and selectivity. Based on our previous studies using fast cyclic square-wave voltammetry (FCSWV) for in vivo dopamine measurements, we have modified this technique to optimize the detection of serotonin in vivo. A series of large amplitude square-shaped potentials was superimposed onto an N-shaped waveform to provide cycling through multiple redox reactions within the N-shaped waveform to increase sensitivity and selectivity to serotonin when combined with a two dimensional voltammogram. N-shaped fast cyclic square-wave voltammetry (N-FCSWV) showed significantly higher sensitivity to serotonin compared to conventional N-FSCV. In addition, N-FCSWV showed better performance than conventional N-shaped FSCV in differentiating serotonin from its major interferents, dopamine and 5-hydroxyindoleascetic acid (5-HIAA). It was also confirmed that the large amplitude of the square waveform did not influence local neuronal activity, and it could monitor electrical stimulation evoked phasic release of serotonin in the rat substantia nigra pars reticulata (SNr) before and after systemic injection of escitalopram (ESCIT, 10 mg/kg i.p.), a serotonin selective reuptake inhibitor.

Single Quantum Dot Tracking Reveals Serotonin Transporter Diffusion Dynamics are Correlated with Cholesterol-Sensitive Threonine 276 Phosphorylation Status in Primary Midbrain Neurons

Article

Full-text available

May 2018

The serotonin transporter protein (SERT) terminates serotonin signaling in the brain by enabling rapid clearance of the neurotransmitter. SERT dysfunction has been associated with a variety of psychiatric disorders, including depression, anxiety, and autism. Visualizing SERT behavior at the single molecule level in endogenous systems remains a challenge. In this study, we utilize quantum dot (QD) single particle tracking to capture SERT dynamics in primary rat midbrain neurons. Membrane microenvironment, specifically membrane cholesterol, plays a key role in SERT regulation and has been found to affect SERT conformational state. We sought to determine how reduced cholesterol content affects both lateral mobility and phosphorylation of conformationally-sensitive threonine 276 (Thr276) in endogenous SERT using two different methods of cholesterol manipulation, statins and methyl-β-cyclodextrin. Both chronic and acute cholesterol depletion increased SERT lateral diffusion, radial displacement along the membrane, mobile fraction, and Thr276 phosphorylation levels. Overall, this work has provided new insights about endogenous neuronal SERT mobility and its associations with membrane cholesterol and SERT phosphorylation status.

Will imaging individual raphe nuclei in males with major depressive disorder enhance diagnostic sensitivity and specificity?

Article

Full-text available

Jan 2018
DEPRESS ANXIETY

Background: Positron emission tomography (PET) studies in major depressive disorder (MDD) have reported higher serotonin 1A (5-HT1A ) autoreceptor binding in the raphe. In males, the difference is so large that it can potentially be used as the first biological marker for MDD. However, the raphe includes several nuclei, which project to different regions of the brain and spinal cord and may be differentially involved in disease. We aimed to identify 5-HT1A differences in individual raphe nuclei using PET in order to determine whether use of subnuclei would provide greater sensitivity and specificity of diagnosing MDD. Methods: We identified individual nuclei using a hybrid set-level technique on an average [11 C]-WAY100635 PET image derived from 52 healthy volunteers (HV). We delineated three nuclei: dorsal raphe nucleus (DRN), median raphe nucleus (MRN), and raphe magnus (RMg). An atlas image of these nuclei was created and nonlinearly warped to each subject (through an associated MRI) in a separate sample of 41 males (25 HV, 16 MDD) who underwent [11 C]-WAY100635 PET. Results: 5-HT1A binding was elevated in DRN in MDD (P < .01), and was not different in the RMg and MRN between groups. Receiver operating characteristic (ROC) curves showed that combining DRN and MRN produces highest sensitivity (94%) and specificity (84%) to identify MDD. Conclusions: In agreement with postmortem studies, we found higher 5-HT1A autoreceptor binding in MDD selectively in the DRN. 5-HT1A autoreceptor binding in the combined DRN and MRN is a better biomarker for MDD than in the raphe as a whole.

Selective Modulation of K+ Channel Kv7.4 Significantly Affects the Excitability of DRN 5-HT Neurons

Article

Full-text available

Dec 2017

The serotonin (5-HT) system originating in the dorsal raphe nucleus (DRN) is implicated in various mood- and emotion-related disorders, such as anxiety, fear and stress. Abnormal activity of DRN 5-HT neurons is the key factor in the development of these disorders. Here, we describe a crucial role for the Kv7.4 potassium channel in modulating DRN 5-HT neuronal excitability. We demonstrate that Kv7.4 is selectively expressed in 5-HT neurons of the DRN. Using selective Kv7.4 opener fasudil and Kv7.4 knock-out mice, we demonstrate that Kv7.4 is a potent modulator of DRN 5-HT neuronal excitability. Furthermore, we demonstrate that the cellular redox signaling mechanism is involved in this 5-HT activation of Kv7.4. The current study suggests a new strategy for treating psychiatric disorders related to altered activity of DRN 5-HT neurons using K+ channel modulators.

Glial Cells

Chapter

Dec 2017

Glia produce trophic factors, ensheath axonal extensions, regulate neurotransmitter and ion concentrations, and remove toxins and debris from the extracellular space of the central nervous system (CNS), maintaining an extracellular milieu that is optimally suited for neuronal function. Consequently, glial functional impairments, as well as physiological reactions of glia to injury, have the potential to induce and/or exacerbate neuronal dysfunction. This mini-review showcases contemporary evidence provoking reformulation of concepts of the interdependence between glia and neurons in modulating final pathways of neuropathologic injury. The chapter commences with a discussion on the role of the various cells in maintaining optimal CNS function both during development and later life-stages, followed by a discussion on their role in mediating neurotoxicity.

The role of the serotonergic system in motor control

Article

Jul 2017

Takashi Kawashima

The serotonergic system in the vertebrate brain is implicated in various behaviors and diseases. Its involvement in motor control has been studied for over half a century, but efforts to build a unified model of its functions have been hampered due to the complexity of serotonergic neuromodulation. This review summarizes the anatomical structure of the serotonergic system, its afferent and efferent connections to other brain regions, and recent insights into the sensorimotor computations in the serotonergic system, and considers future research directions into the roles of serotonergic system in motor control.

Depressive-like neurochemical and behavioral markers of Parkinson’s disease after 6-OHDA administered unilaterally to the rat medial forebrain bundle

Article

Jun 2017

Background: Although Parkinson's disease (PD) is characterized by progressive neurodegeneration of multiple neurotransmitter systems, 6-hydroxydopamine (6-OHDA) as a model substance is mainly used to selectively damage the nigrostriatal dopaminergic neurons and induce parkinsonian-like motor disturbances in rats. We hypothesized that high doses of this neurotoxin affecting other monoaminergic systems may also evoke the depressive-like behavior. Methods: The impact of 6-OHDA (8, 12, 16μg/4μl) administered unilaterally into the medial forebrain bundle on the sucrose solution intake (a measure of anhedonia) and on the tissue levels of noradrenaline (NA), dopamine (DA) and serotonin (5-HT) in the striatum (STR), substantia nigra (SN), prefrontal cortex (PFC) and hippocampus (HIP) was examined in rats pretreated or non-pretreated with desipramine. Results: The highest dose of 6-OHDA reduced the preference for 3% sucrose solution both in rats without and with desipramine pretreatment. All used doses of 6-OHDA dramatically decreased DA content in the studied brain structures on the ipsilateral side. NA levels were severely decreased in the ipsilateral STR, HIP and PFC of rats non-pretreated with desipramine and to a much lesser extent in those pretreated with desipramine. In the SN, moderate decreases in NA level were found both in rats pretreated and non-pretreated with desipramine. Higher doses of 6-OHDA reduced 5-HT content in the ipsilateral STR, HIP and PFC, but not in the SN, only in rats non-pretreated with desipramine. Conclusions: Administration of the highest dose of 6-OHDA without desipramine pretreatment evoked neurochemical and behavioral changes resembling the advanced PD with coexisting depression.

Anatomy of the central serotoninergic projection systems

Article

Jan 2000

Serotoninergic neurons are integral parts of central and/or peripheral nervous networks in diverse forms of invertebrates and vertebrates (Parent 1981a,b), suggesting that these neurons provide animals - across phylogeny - with capacities essential for adapting to changing internal and/or external demands. Clues as to their functional role(s) may already be gained from their morphology: giant metacerebral serotoninergic neurons in molluscs have abundantly collateralized axons (Cottrell 1977) as do some of the large multipolar, extensively ramifying serotoninergic neurons of the mammalian raphe nuclei/ extraraphe reticular 5-HT cell groups (Waterhouse et al. 1986; Vertes 1991; Van Boeckstaele et al. 1993; Vertes and Kocsis 1994; Holmes et al. 1994), implying that they innervate multiple target networks along the neuraxis. This enables them to coordinate and harmonize activities (or response properties) in diverse networks with state-dependent determinants such as the prevailing level of central motor tone, (somato)-sensory processing and autonomic regulation across the sleep/wake cycle (Jacobs and Fornal 1993). By way of abundantly collateralizing, a limited number of neurons are capable of modulating electrical activity and afferent input responsivity in multiple targets in a coordinate fashion. Therefore, certain serotoninergic neurons of the brainstem represent archetypical reticular-type multipolar neurons which resemble the Golgi type 1 neurons of the brainstem reticular core described by Scheibel and Scheibel(1958).

Strategies to Optimize the Antidepressant Action of Selective Serotonin Reuptake Inhibitors

Chapter

Jan 1997

It is beyond doubt that major depression can be treated by selectively manipulating the function of different brain neurotransmitters (1,2). Yet, of the many brain neuronal systems, the serotonin (5-hydroxytryptamine, 5-HT) system is the most common neurobiological target for such treatments. Tricyclic antidepressants (TCAs) act on 5-HT and noradrenergic (NE) neurons by inhibiting, with different potencies, transmitter reuptake (3,4), and monoamine (MAO) oxidase inhibitors (MAOIs) increase 5-HT and NE transmission by preventing their metabolism. Yet, it was not until the advent of the selective serotonin reuptake inhibitors (SSRIs) that the antidepressant potential of 5-HT transporter blockade was fully appreciated (5,6). The clinically useful SSRIs are chemically dissimilar, but share the property of selectively inhibiting the 5-HT reuptake process (Fig. 1) Unlike TCAs, the SSRIs display little affinity for aminergic receptors (7) and therefore lack the severe side effects associated with the use of the former agents. This results in both an improved quality of life for the patients and greater treatment compliance, which is compromised in some instances by the use of TCAs. It is generally recognized that the antidepressant efficacy of SSRIs is comparable to that of TCAs, although several studies have shown that the latter are more effective in severely depressed inpatients (5,6,8,9).

Nicotine and Stimulatory Effects on 5-HT DRN Neurons

Chapter

Dec 2016

A large number of behaviors and brain functions are modulated by nicotine through stimulation of presynaptic and, less frequently, postsynaptic nicotinic acetylcholine receptors (nAChRs). Clinical and experimental studies suggested that many of nicotine's behavioral effects are mediated by the brain serotonergic (5-HT) system. Dorsal raphe nucleus (DRN) neurons give most of the forebrain 5-HT innervation. Brain slice experiments indicated that nicotine increases the firing activity of 60–80% of 5-HT DRN neurons. The stimulatory effects of nicotine are direct, dependent on α7 and α4β2 somatodendritic nAChRs, and indirect, dependent on presynaptic nAChRs located on glutamatergic (α4β2 nAChRs) and noradrenergic (α7 nAChRs) terminals in the DRN. Microdialysis experiments indicated that systemic nicotine increases 5-HT release in a large number of cerebral areas. Likewise, it was shown that most of nicotine's behavioral effects are mediated by stimulation of various subtypes of 5-HT receptors.

Neuroanatomie und Neurophysiologie des zentralen 5-HT-Systems

Chapter

Jan 1991

Hans Georg Baumgarten

Fortschritte in Methoden zur experimentellen und klinischen Erforschung verschiedenster Aspekte serotonerger Mechanismen haben zur Modifizierung früherer Konzepte des zentralen Serotoninsystems als eines vorwiegend „trophotropen“ Neurotransmitters geführt und klargestellt, daß die modulierenden Eigenschaften dieses expansiven und verbindungsreichen Neuronensystems in fast allen zentral regulierten Leistungen nachweisbar sind (Kognition, Lernen, Engrammbildung, Vigilanzsteuerung; Affekt-, Stimmungs- und Impulskontrolle; Sexual- und Sozialverhalten, Triebbefriedigungsverhalten, Alarmqualität von fremden Wahrnehmungsmustern, Schwellenempfindlichkeit für sensorische Stimulusmuster und für motorische Reflexreagibilität; Dynamik des motorischen Antriebspegels; Kreislauf- und Atemregulation; zirkadiane Organisation von neuroendokrinen Sekretionsprofilen, von Temperatur-, Aktivitäts- und Stimmungszyklen). Zur Sicherstellung so weitreichender Einflüsse auf kortexabhängige komplexe Funktionen hat sich das Serotoninsystem beim Primaten an die Evolution limbischer und sensorischer Kortizes angepaßt.

A study of subcortical afferents to the hippocampal formation in the rat

Article

Full-text available

Feb 1979
NEUROSCIENCE

The distribution of neurons in diencephalon and brainstem which project upon the hippocampal formation has been analyzed in adult rats by the injection of horseradish peroxidase into different parts of the hippocampus and dentate gyrus and the related retrohippocampal structures, including the subicular complex and the entorhinal cortex. Any large injection of horseradish peroxidase inlo the hippocampal region results in the retrograde labeling of some neurons in each of the following structures: in the thalamus—the nucleus reuniens, the parataenial and paraventricular nuclei, the anterodorsal and antermedial nuclei, and the laterodorsal and lateral posterior nuclei; in the hypothalamus, septum and preoptic region—the medial septal nucleus and the diagonal band of Broca, the substantia innominata, the lateral preoptic area, the magnocellular preoptic nucleus, and the anterior amygdaloid area, the dorsomedial hypothalamic nucleus, the lateral and posterior hypothalamic areas, the ventral premammillary nucleus, the supramammillary region, and parts of the tuberomammillary and lateral and medial mammillary nuclei: in the brainstem: the ventral tegmental area, the substantia nigra, the interpeduncular and interfascicular nuclei, the dorsal and median nuclei of the raphe, the dorsal and laterodorsal tegmental nuclei, the locus coeruleus, the central gray, and certain of the tegmental reticular fields.It is clear that these afferents to the hippocampal formation do not comprise a single, homogeneous system, and that their terminations within the hippocampal region are not restricted to a particular topographic level (i.e. septal, intermediate or temporal).

WITHDRAWN: References

Chapter

Jan 1982

Evidence for the existence of monoamine containing neurons in the central nervous system. IV. Distribution of monoamine nerve terminals in the central nervous system

Article

Jan 1965

Kjell Fuxe

A stereotaxic atlas of the forebrain and lower parts of the brain stem

Article

Jan 1974

Divergent axon collaterals of rat locu Demonstration by a fluorescent double labeling technique

Article

Sep 1981
BRAIN RES

The distribution of the noradrenaline-containing neurons of the rat locus coeruleus has been investigated with retrograde labeling techniques using two different fluorescent tracers. Injections were placed in the prefrontal cortex, the striatum, the thalamus, the hippocampus, the cerebellar cortex and the lumbar spinal cord.No evidence for locus coeruleus projections to the striatum was found. Injections in the cortex, thalamus and hippocampus revealed not only ipsilateral but also contralateral labeling of cells in the locus coeruleus. Following unilateral or bilateral homo- or heterotopic injections of the two tracers several cells of the locus coeruleus were double labeled. Combined injections of the two fluorophores in any of these forebrain areas and in the spinal cord also produced double labeled cells. The majority of double labeled cells was located in an area between the ventral and the dorsal parts of the locus coeruleus.These results indicate that individual neurons of the locus coeruleus have the possibility to influence adrenergic receptors at remote areas in the central nervous system.

Dahlstrom A, Fuxe K. Evidence for existence of monoamine-containing neurons in central nervous system. I. Demonstration of monoamines in cell bodies of brain stem neurons. Acta Physiol Scand 232(Suppl): 1-55

Article

Nov 1963
Acta Physiol Scand Suppl

An autoradiographic analysis of the differential ascending projections of the dorsal and median raphe nuclei in the rat

Article

Jun 1978
J COMP NEUROL

The differential projections from the dorsal raphe and median raphe nuclei of the midbrain were autoradiographically traced in the rat brain after 3H-proline micro-injections. Six ascending fiber tracts were identified, the dorsal raphe nucleus being the sole source of four tracts and sharing one with the median raphe nucleus. The tracts can be classified as those lying within the medial forebrain bundle (dorsal raphe forebrain tract and the median raphe forebrain tract) and those lying entirely outside (dorsal raphe arcuate tract, dorsal raphe periventricular tract, dorsal raphe cortical tract, and raphe medial tract). The dorsal raphe forebrain tract lies in the ventrolateral aspect of the medial forebrain bundle (MFB) and projects mainly to lateral forebrain areas (e.g., basal ganglion, amygdala, and the pyriform cortex). The median raphe forebrain tract lies in the ventromedial aspect of the MFB and projects to medial forebrain areas (e.g., cingulate cortex, medial septum, and hippocampus). The dorsal raphe cortical tract lies ventrolaterally to the medial longitudinal fasciculus and projects to the caudate-putamen and the parieto-temporal cortex. The dorsal raphe periventricular tract lies immediately below the midbrain aqueduct and projects rostrally to the periventricular region of the thalamus and hypothalamus. The dorsal raphe arcuate tract curves laterally from the dorsal raphe nucleus to reach the ventrolateral edge of the midbrain and projects to ventrolateral geniculate body nuclei and the hypothalamic suprachiasmatic nuclei. Finally, the raphe medial tract receives fibers from both the median and dorsal raphe nuclei and runs ventrally between the fasciculus retroflexus and projects to the interpeduncular nucleus and the midline mammillary body. Further studies were done to test whether the fiber tracts travelling in the MFB contained 5-HT. Unilateral (left) injections of 5,7-dihydroxytryptamine (5 μgm/400 nl) 18 days before midbrain raphe microinjections of 3H-proline produced a reduction in the grain concentrations in all the ascending fibers within the MFB. Furthermore, pharmacological and behavioural evidence was obtained to show that the 5-HT system had been unilaterally damaged; these animals displayed preferential ipsilateral turning in a rotameter which was strongly reversed to contralateral turning after 5-hydroxytryptophan administration. The results show that DR and MR nuclei have numerous ascending projections whose axons contain the transmitter 5-HT. The results agree with the neuroanatomical distribution of the 5-HT system previously determined biochemically, histochemically, and neurophysiologically. The midbrain serotonin system seems to be organized by a series of fiber pathways. The fast transport rate in these fibers was found to be about 108 mm/day.

Subcortical afferent connections of the amygdala in the monkey

Article

Apr 1980
J COMP NEUROL

William R. Mehler

The cells of origin of the afferent connection of the amygdala in the rhesus and squirrel monkeys were analyzed by means of the retrograde axonal transport of the enzyme horseradish peroxidase (HRP) injected into various quadrants of the amygdala. Tabulation of the relative numbers of HRP labeled cells found in each brain section through various nuclei in a series of injected brains revealed several patterns of ipsilateral subcortical nuclear connections with the amygdala. The dorsal thalamic nuclei most consistently exhibiting large numbers of labeled cells in all experiments were the ipsilateral halves of the midline nucleus paraventricularis thalami and both the parvo‐ and magnocellular parts of the nucleus subparafascicularis. All of the subdivisions of the midline nucleus centralis complex (Olszewski, '52) exhibit HRP labeled cells in most cases. A cell group corresponding to the nucleus reuniens ventralis (Kuhlenbeck, '54) and one identified as the nucleus interventralis of Aronson and Papez ('34) also were found to contain labeled cells. The largest populations of HRP labeled cells in the hypothalamus appear chiefly in the middle and posterior parts of the ipsilateral, lateral hypothalamic area in all cases and densely in the ventromedial hypothalamic nucleus only in cases with medial amygdalar quadrant injections. Scattered HRP positive cells frequently can be found in the supramammillary and dorsomedial nuclei and the posterior hypothalamic area. A limited but consistent number of labeled cells can be located in Tsai's ventral tegmental area. In the midbrain some labeled cells can be demonstrated in the rostral and caudal subdivisions of the nucleus linearis, and scattered groups of HRP positive cells can be localized in the dorsal raphe nucleus, chiefly on the side ipsilateral to the injection. The most conspicuous subdiencephalic source of amygdalar afferent connections found in the present study is the pars lateralis of the nucleus parabrachialis located in the dorsolateral pontine tegmentum. Cells in this nucleus are heavily labeled in all cases, while only a few HRP positive cells are occasionally found in the pars medialis. A few labeled cells can be differentiated from pigmented cells in the locus coeruleus, but the labeling of cells scattered throughout the caudal half of the nucleus of the tractus solitarius and in a paravagal cell group is accepted with reservations because both cell groups frequently display endogenously pigmented cells in normal, non‐injected monkeys.

Geyer MA, Puerto A, Menkes DB, Segal DS, Mandell AJ. Behavioral studies following lesions of the mesolimbic and mesostriatal serotonergic pathways. Brain Res 106: 257-269

Article

May 1976
BRAIN RES

The behavior of rats with selective lesions of either the dorsal (B7), median (B8), or lateral (B9) raphe nuclei was compared to that of sham-lesioned controls in a variety of experimental situations. As described previously17, the extent of damage to the midbrain raphe nuclei was determined by fluorescence histochemistry, and the tryptophan hydroxylase and tyrosine hydroxylase activities of 6 forebrain regions were measured for each rat. None of the lesions affected tyrosine hydroxylase activity. Lesions of B7, which reduced tryptophan hydroxylase in the striatum, thalamus, cortex, and hypothalamus, had no significant effect on any of the behavioral measures. Lesions of B9, although twice as large, neither reduced forebrain tryptophan hydroxylase significantly nor affected any of the behavioral variables. However, B8 lesions, which reduced hippocampal, septal, cortical, and hypothalamic tryptophan hydroxylase, had behavioral effects similar to those reported after combined raphe lesions or parachlorophenylalanine. Median raphe-lesioned rats were hyperactive when placed in a novel environment and throughout the dark phase of the light/dark cycle. With respect to locomotor activity, B8-lesioned rats were also hyper-responsive to amphetamine. When placed in a stabilimeter and subjected to repeated air puff stimuli, rats with B8 lesions exhibited larger startle responses. Furthermore, only B8-lesioned animals perseverated when given two unreinforced trials in a Y-maze. All these histologic, biochemical, and behavioral variables were assessed individually for all 39 animals, and a multivariate correlational analysis incorporating the data of this and the preceding paper17 is presented here. These experiments suggest that the mesolimbic serotonergic pathway originating in B8 subserves some of the inhibition necessary to dampen behavioral responsivity.

Monoamine innervation of the forebrain: Collateralization

Article

Jul 1982
BRAIN RES BULL

The forebrain is characterized by a dense, localized dopamine (DA) innervation pattern, a diffuse, widespread norepinephrine (NE) innervation pattern, and a serotonin (5-HT) innervation intermediate between the DA and NE patterns. These innervation patterns have implied that basic differences exist in the way DA, NE and 5-HT axons collateralize to different brain structures; that is, DA axons are thought to be poorly collateralized and NE and 5-HT axons are presumed to be more highly collateralized. In the present study, we used injections of retrograde labeling fluorescent dyes into various forebrain regions in order to determine axonal branching patterns from nuclei that contain DA, NE and 5-HT neurons, namely the substantia nigra-ventral tegmental area (SN-VTA), locus coeruleus (LC) and raphe nuclei (DR-MR). The results suggest that at least two subpopulations of neurons can be defined in each monoamine nucleus with respect to the way their axons collateralize. Each area contains a centrally located nuclear area with highly collateralized neurons, and more peripherally situated areas with less highly collateralized neurons. Thus, previous suppositions of the branching of monoamine axons must be revised to account for the existence of cells exhibiting totally different Collateralization patterns within each monoamine nucleus.

Histologic and enzymatic studies of the mesolimbic and mesostriatal serotonergic pathways

Article

May 1976
BRAIN RES

Selective lesions of the dorsal (B7), median (B8), or lateral (B9) raphe nuclei were made stereotaxically in male rats 4 weeks before sacrifice. The extent of damage to each raphe nucleus was quantified histologically by means of a simplified formaldehyde histochemical method for visualization of serotonin in cryostat sections. A detailed mapping of the distribution of the yellow-fluorescent raphe perikarya provided the basis for quantification. Tryptophan hydroxylase activity was measured in 6 forebrain regions from each animal, and the results were correlated with the per cen damage to each raphe nucleus. Tyrosine hydroxylase was also assayed in 5 of these regions; it was not significantly affected by any of the raphe lesions. Dorsal raphe lesions reduced tryptophan hydroxylase activity in the striatum, thalamus, cortex, and hypothalamus, but not in the septal nuclei or hippocampus. Damage to B8 resulted in decrements in this serotonergic enzyme in the septal nuclei, hippocampus, cortex, and hypothalamus, but not in the striatum or thalamus. Lesions of the scattered B9 cells had no significant effect on enzyme activity in any region examined. These data suggest that the dorsal and median raphe nuclei provide two distint though perhaps overlapping serotonergic systems innervating different parts of the forebrain: a mesostriatal pathway originating in B7 and a mesolimbic system derived from B8. Behavioral studies on the animals, which are presented in a companion paper9, indicated that damage to the median nucleus is responsible for many of the behavioral effects previously reported after combined lesions of both major raphe nuclei.

Distribution of serotonin-immunoreactivity in the central nervous system of the rat—Cell bodies and terminals

Article

Feb 1981
NEUROSCIENCE

Harry W M Steinbusch

The localization and distribution of serotonin (5-hydroxytryptamine, 5-HT) has been studied with the indirect immunofluorescence technique using a highly specific and well-characterized antibody to 5-HT. In neuron systems 5-HT was found to be primarily present with a distribution similar to that observed in basic mappings carried out with the formaldehyde-induced fluorescence method. In addition to the nine areas originally described, several other areas in the mesencephalon and rhombencephalon appeared to contain widely distributed 5-HT-positive perikarya. In the median eminence 5-HT fluorescent mast cells could be visualized. No 5-HT-positive nerve cell bodies could be observed either in the telencephalon or diencephalon.

A comparison of the retrograde tracer properties of [125I] wheat germ agglutinin (WGA) with HRP after injection into the corpus callosum

Article

Mar 1982
BRAIN RES

The autoradiographic demonstration of axonal connections in the CNS

Article

Feb 1972
BRAIN RES

Stereotaxic Mapping of the Monoamine Pathways in the Rat Brain*

Article

Dec 1971
Acta Physiol Scand Suppl

Urban Ungerstedt

The ascending monoamine pathways in the rat brain are demonstrated by the pile up of fluorescent material occurring in the axons after various types of lesions. The anatomy of the pathways is outlined in drawings of frontal sections of the brain and the origin and termination of several pathways is determined by studying the anterograde and retrograde degeneration occurring after well localised lesions. It is possible to separate the ascending NA pathways into a dorsal and a ventral bundle of axons. The dorsal bundle innervates the cortex and the hippocampus and the ventral bundle supplies NA nerve terminals to the medulla, the pons, the mesencephalon and the diencephalon. The dorsal bundle is found to originate in the locus coeruleus. Lesions of this nucleus abolish the nerve terminals in all cortical areas and in several other areas of the brain indicating a unique role for the locus coeruleus in influencing the activity of the entire brain. The 5-HT pathways have a distribution similar to the ventral NA pathyway. The course of the nigro-striatal and the meso-limbic DA pathways is presented in detail.

Differential projections of the nucleus Raphe dorsalis and nucleus Raphe centralis as revealed by autoradiography

Article

Mar 1975
BRAIN RES

Differential projections of neurons within the dorsal raphe nucleus of the rat: A horseradish peroxidase (HRP) study

Article

Jun 1978
BRAIN RES

Application of coupled oxidation reaction to electron microscopic demonstration of horseradish peroxidase: cobalt-glucose oxidase method

Article

Nov 1979
BRAIN RES

Raphe projections to the locus coeruleus in the rat

Article

Jul 1979
BRAIN RES BULL

Afferent projections to the locus coeruleus from the various raphe nuclei, particularly of the midbrain (nuclei raphe dorsalis and medianus) and pons (nuclei raphe pontis and magnus), have been studied in the rat by retrograde transport methods using horseradish peroxidase (HRP). The locus coeruleus, in both its dorsomedial and ventrolateral divisions, and in its various anterior-posterior components, were injected with 0.05 microliters of horseradish peroxidase following which various structures of the brainstem, particularly the raphe nuclei, were examined for HRP reactive cells. It was found that injections in most components of the locus coeruleus were associated with HRP positive cells in varying degrees of density in the nuclei raphe dorsalis, medianus, pontis, and magnus, with considerably sparser labelling in the anterior aspects of the medullary raphe nuclei pallidus and obscurus. Labelled cells were also seen in the nuclei of the solitary tract, contralateral locus coeruleus. lateral reticular areas of the pons and midbrain, nuclei pontis oralis and caudalis, vestibular nuclei, mesencephalic nucleus of the trigeminal nerve, fastigial nuclei of cerebellum and medial parabrachial nuclei. These data, showing widespread innervation of the locus coeruleus from all raphe nuclei, as well as many other brainstem areas, in the rat support the general view of heavy innervation of the locus coeruleus from both extra-raphe and raphe nulcei. These latter raphe projections, probably serotonergic in nature, provide anatomical support for the various experiments indicating considerable regulation of locus coeruleus activities, such as phasic events of REM sleep, among other, by widespread projections from most raphe nuclei was well as several other regions of the brainstem.

Serotonin and Substance P Coexist in Neurons of the Rat's Central Nervous System

Article

Apr 1978

5-Hydroxytryptamine (serotonin)-containing neurons in the rat's medullary raphe and interfascicularis hypoglossi cell groups were identified by means of autoradiography following prolonged intraventricular administration of 5-hydroxy[(3)H]tryptamine, fluorescence histochemistry for the demonstration of endogenous 5-hydroxytryptamine, and microspectrofluorimetric analysis of excitation and emission spectra. Immunocytochemical methods (the unlabeled primary antibody-peroxidase antiperoxidase and indirect immunofluorescence methods) were applied with antisera to substance P in order to localize immunoreactivity in these medullary neurons. It was demonstrated that the raphe nuclei and the interfascicularis hypoglossi nucleus are heterogeneous cell groups that contain: (i) Neurons that display both an uptake-storage capacity for 5-hydroxy[(3)H]tryptamine and a formaldehyde-induced fluorescence with spectral characteristics identical to those of the 5-hydroxytryptamine fluorophor. These cells exhibit high to low fluorescence intensities without detectable substance P-like immunoreactivity. (ii) Neurons with various 5-hydroxytryptamine fluorescence intensities and intense to low degrees of substance P-like immunoreactivity. (iii) Neurons with various degrees of substance P-like immunoreactivity without detectable 5-hydroxytryptamine fluorescence or 5-hydroxy[(3)H]tryptamine uptake and storage capacity. These results indicate that some neurons contain high or low levels of only 5-hydroxytryptamine or substance P, whereas other neurons contain both 5-hydroxytryptamine and substance P in various proportions. The present findings demonstrate the presence of two putative transmitters, a biogenic amine and a polypeptide, within the same neuron in the mammalian central nervous system.

Excitation of caudate-putamen neurons following stimulation of dorsal raphe nucleus in the rat

Article

Nov 1979
BRAIN RES

The cerebellar projection from the raphe nuclei in the cat as studied with the method of retrograde transport of horseradish peroxidase

Article

Jan 1978

Following injections of horseradish peroxidase (HRP) in the cerebellar cortex and nuclei of the cat, the distribution of labeled cells in the raphe nuclei was mapped. The findings confirm those made previously in studies of retrograde cell degeneration following cerebellar ablations (Brodal et al., 1960a), and in addition reveal new details in the projection of the raphe nuclei onto the cerebellar cortex and nuclei. All the raphe nuclei except nucleus linearis intermedius and nucleus linearis rostralis project onto the cerebellar cortex. The nuclei raphe obscurus and pontis contribute the greatest number of afferents to the cerebellum. With the exception of lobule VI which probably is the recipient of a weak projection, all parts of the cerebellar cortex receive afferents from the raphe nuclei. The heaviest projection is to the vermis of lobules VIIA and X, and to crus II. The afferents to the cerebellar nuclei are few in number (Tables 2–6). The observations indicate that each raphe neuron probably projects to more than one terminal site in the cerebellum. The findings are discussed with reference to other efferent and afferent studies of the raphe nuclei. All these studies indicate that the raphe nuclei have widespread efferent and afferent connections, making them capable to participate in a variety of regulatory functions.

Afferent projections to the rat locus coeruleus as determined by a retrograde tracing technique

Article

Mar 1978

Afferent connections to the rat locus coeruleus (LC), which contains exclusively noradrenergic neurons, have been traced using the technique of retrograde transport of horseradish peroxidase (HRP). In order to ensure accurate placement of adequate amounts of HRP in the LC, a microiontophoretic delivery technique coupled with single cell recording was employed. The use of electro physiological "landmarks" as aids in placing the injections is described. Following HRP injections into the LC, forebrain structures containing labelled neurons included the insular cortex, the central nucleus of the amygdala, the medial, lateral and magnocellular preoptic areas, the bed nucleus of the stria terminalis, and the dorsomedial, paraventricular and lateral hypothalamic areas. In the brainstem reactive neurons were observed in the central grey substance, the reticular formation, the raphe, vestibular, solitary tract and lateral reticular nuclei. In particular, the areas of catecholamine cell groups A1, A2 and A5 appeared to contain many reactive cells. Labelled neurons were also observed in the fastigial nuclei and in the marginal zones of the dorsal horns of the spinal cord. This pattern of afferent innervation supports suggestions for a role for the LC in behavioral arousal mechanisms and autonomic regulation.

The raphe nuclei of the cat brain stem: A topographical atlas of their efferent projections as revealed by autoradiography

Article

Oct 1976
BRAIN RES

Stereotaxic injections of [14C]leucine were made in nulei raphe centralis superior, raphe dorsalis, raphe magnus and raphe pontis of the cat. The organization of the regional connections was outlined in a stereotaxic atlas using the autoradiographic tracing method: the majority of the ascending pathways from the rostral raphe nuclei are directed mainly through a ventrolateral bundle via the ventral tegmental area of Tsai, with some lateral extensions to the substantia nigra, and then through the fields of Forel and the zona incerta. More rostrally the fibers are joined to the medial forebrain bundle through the hypothalamic region up to the preoptic area or the diagonal band of Broca. Multiple divisions leave this tract towards the epithalamic or the intralaminar thalamic nuclei, the stria terminalis, the septum, the capsula interna and the ansa lenticularis. The bulk of the rostral projections terminates in the frontal lobe, while some labeling is scarcely distributed throughout the rest of the neocortex. The projections of nucleus (n.) raphe centralis superior are specifically associated with the n. interpeduncularis, the mammillary bodies and the hippocampal formation while the n. raphe dorsalis innervates selectively the lateral geniculate bodies, striatus, piriform lobes, olfactory bulb and amygdala. The rest of the ascending fibers form the centrolateral or the dorsal ascending tracts radiating either in the reticular mesencephalic formation or in the periventricular gray matter. On the contrary there are heavy descending projections from n. raphe centralis superior which distribute to the main nuclei of the brain stem, the central gray matter and the cerebellum. The ascending projections form the caudal raphe nuclei are much less dense. They disseminate mainly in the colliculus superior, the pretectum, the nucleus of the posterior commissure, the preoculomotor complex and the intralaminar nuclei of the thalamus. From n. raphe pontis, a dense labeling is selectively localized at the n. paraventricularis hypothalami with some rostral extensions to limbic areas. Diffuse caudal and rostral projections from both nuclei are observed in the mesencephalic, pontobulbar reticular formation and the cerebellum. The main differences come from the specific localization of their descending bulbospinal tracts inside the lateroventral funiculus of the spinal cervical cord.

Differential efferent connections of the brain stem to the hippocampus in the cat

Article

Feb 1977
BRAIN RES

Behavioral effects of selective midbrain raphe lesions in the rat

Article

Jun 1975
BRAIN RES

Lesions were produced in the median (n = 8), dorsal (n = 7) or both (n = 7) midbrain raphe nuclei and their effects on behavior (days 16-54 postoperatively) compared to that of controls (n = 9). In addition, forebrain 5-hydroxytryptamine (5-HT) concentration were determined. Only the median and combined lesion groups showed increased running wheel and open field activity, as well as enhanced reactivity to novel stimuli and environmental change. None of the lesion groups, however, showed changes in home cage activity on postoperative day 21. Although all lesion groups were deficient in the acquisition and retention of one-way avoidance, the deficits were of a greater magnitude in the median and combined lesion groups. The latter two groups, furthermore, were impaired in forced extinction of the one-way avoidance response, but only the combined lesion group evidenced facilitation of two-way avoidance acquistion. Thus, in contrast to the effects of median or combined raphe lesions, lesions in the dorsal raphe nucleus affected few of the behavioral parameters studied. These results suggest that the dorsal raphe nucleus plays a different behavioral role than the median raphe nucleus. The median nucleus appears to be involved in the regulation of activity level, the reaction to novelty and environmental change, and the response to aversive stimuli. Possible mechanisms for the observed behavioral changes are discussed, as well as their apparent similarity to the effects of other mesencephalic and limbic lesions. Lastly, the median, dorsal and combined raphe lesions lowered forebraine 5-HT but 26, 65, and 77%, respectively, versus controls. These reductions differed significantly from each other, and with previously reported data indicate that the dorsal raphe nucleus in the principal origin of forebrain 5-HT. It is suggested, furthermore, that the behavioral effects of midbrain raphe lesions are not due primarily to their associated reduction in forebrain 5-HT.

A raphe projection to cat cerebellar cortex

Article

Nov 1975
BRAIN RES

Anatomical and electrophysiological identification of a projection from the mesencephalic raphe to the caudate-putamen in the rat

Article

Nov 1975
BRAIN RES

Trigeminocerebellar, trigeminotectal, and trigeminothalamic projections: A double retrograde axonal tracing study in the mouse

Article

Jul 1985

Dennis Steindler

Double retrograde axonal tracing experiments were carried out in order to reveal potential patterns of divergence in axonal projections from the two major sensory nuclei of the mouse brainstem trigeminal complex: the principal sensory and spinal trigeminal nuclei (oralis, interpolaris, and caudalis divisions). The tracers wheat germ agglutinin, N‐[acetyl‐ ³ H] and horseradish peroxidase were used in paired injection strategies within portions of the cerebellum, superior colliculus, and thalamic ventrobasal complex and/or posterior group of adult ICR white mice. Trigeminal neurons with projections to tactile areas of the cerebellar cortex or underlying deep cerebellar nuclei were found scattered throughout the principal sensory nucleus and interpolaris division, and mainly in dorsal regions of the oralis division of the spinal trigeminal nucleus. Injections of either tracer which involved lateral portions of the rostral half of the superior colliculus labeled trigeminotectal neurons mainly in the contralateral interpolaris division, ventral half of the oralis division, and a ventral region of the principal sensory nucleus near the oralis border. Fewer trigeminotectal neurons were found scattered throughout the principal sensory nucleus and the magnocellular layer of the caudalis divisions, although an occasional labeled neuron was also found in the marginal layer. Contralaterally projecting trigeminothalamic neurons were observed throughout the principal sensory nucleus, interpolaris division, and within the marginal and magnocellular layers of caudalis. Double‐labeled neurons were observed only after paired injections of the tracers in the thalamus and ipsilateral superior colliculus, and they were found within the caudoventral portion of the principal sensory nucleus near the oralis border, throughout the interpolaris division, within the magnocellular layer of caudalis, and only a few double‐labeled neurons were also found within the marginal layer. After such injections. 50% of the labeled tectum‐projecting neurons in the principal sensory nucleus, 64% in the interpolaris division, and 57% in the caudalis division are branched neurons which have collateralized projections to both the superior colliculus and thalamus. These projections, which have not been described before, appear to arise from more than one class of projection neuron which is differentially distributed within different regions of the trigeminus.

Corticocortical and Collateral Thalamocortical Connections of Postcentral Somatosensory Cortical Areas in Squirrel Monkeys: A Double-Labeling Study with Radiolabeled Wheatgerm Agglutinin and Wheatgerm Agglutinin Conjugated to Horseradish Peroxidase

Article

Feb 1985
Somatosens Res

Experiments were performed to determine whether thalamic neurons send collaterals to more than one subdivision of postcentral somatosensory cortex, and to evaluate the fidelity of somatotopic connections between cortical representations of the body surface. After microelectrode recordings identified the locations of the representations of the fingertips along the border of area 3b with area 3a and along the border of area 1 with area 2, one tracer, tritiated N-acetyl wheatgerm agglutinin ([3H]WGA), was injected into the rostral representation at the 3a-3b border, and another, wheatgerm agglutinin conjugated to horseradish peroxidase (WGA:HRP), was injected into the caudal representation at the 1-2 border. The results indicate that three thalamic regions are interconnected with both border zones. Regions of overlapping retrograde and anterograde label included the ventromedial portion of the lateral division of the ventroposterior nucleus (VPL), which is known to represent the digits; the medial extent of a dorsal capping zone of VPL that we term the ventroposterior superior nucleus (VPS); and the anterior pulvinar (Pa). Cells labeled with one or the other tracer, as well as cells labeled with both tracers, were found in VPL, VPS, and Pa. Our procedures necessarily underestimated the numbers of cells projecting to both rostral and caudal injection zones, but at least 15-20% of VPL and 20-40% of VPS neurons that were autoradiographically labeled appeared to have such collaterals. While only a few cells in Pa were autoradiographically labeled from injections at the 3a-3b border, the presence of several double-labeled cells suggests that at least some Pa neurons project to both injected zones. By relating the present results to those from studies where connections were determined for single architectonic fields, we conclude that many neurons in VP project to both areas 3b and 1, and that many neurons in VPS project to both areas 3a and 2. In addition, both areas 3a and 2 appear to have interconnections with Pa. Connections of Pa with area 2 have only recently been reported (Pons and Kaas, in press), and connections of Pa with area 3a have not been described. The results imply that, in part, the same information from cutaneous receptors is relayed to areas 3b and 1, and the same information from deep receptors is relayed to areas 3a and 2.(ABSTRACT TRUNCATED AT 400 WORDS)

Differential behavioral and neurochemical effects following lesions of the dorsal or median raphe nuclei in rats

Article

Nov 1974
BRAIN RES

Locomotor activity and regional forebrain levels of serotonin (5-HT) were measured following the placement of electrolytic lesions in either the dorsal or median raphe nucleus of adult male rats. In the first two experiments, control lesions were placed in the brachium conjunctivum, and in the third experiment, a sham lesion group served as control. Median lesions significantly increased locomotor actiivity as measured in tilt cages, by 250–300% on the second day post-lesion, and this elevation stabilized at approximately 100% above control levels on day 9 post-lesion. There were no statistically significant differences in the amount of locomotor activity in the dorsal, brachium or sham lesioned groups on any post-lesion day. When the amount of 5-HT depletion was measured 5 days post-lesion, it was found that the dorsal (D) and median (M) lesions produced similar reductions in cerebral cortex (D — 40%; M — 31%); hypothalamus (D — 54%; M — 58%) and striatum (D — 50%; M — 29%). However, the effects of the two lesions were markedly different in the hippocampus. The dorsal lesion produced a non-significant 10% reduction in hippocampal 5-HT level, while the median lesion caused an 82% reduction. On the basis of these data it is hypothesized that a reduction in hippocampal 5-HT may account for the increased activity in the median lesioned group.

Regional 5–hydroxytryptamine following selective midbrain raphe lesion in the rat

Article

Oct 1974
BRAIN RES

Lesions were produced in either the dorsal or median raphe nucleus and regional 5-hydroxytryptamine (5-HT), 5-hydroxyindole acetic acid (5-HIAA), and norepinephrine (NE) determined 26–30 days post-operatively. Only lesions in the dorsal raphe nucleus produced a fall (54%) in striatal 5-HT, while only lesions in the median raphe nucleus reduced (62%) hippocampal 5-HT. Reductions in the 5-HT and 5-HIAA contents of the remaining portion of the telencephalon and of the diencephalon following dorsal raphe lesions were twice as great as after median raphe lesions. The lowered 5-HT concentration (50%) of the telecephalon (excluding hippocampus and striatum) after dorsal raphe lesions, furthermore, was twice as large as that of the diecephalon (24%). Only dorsal raphe lesions produced a fall in brain stem 5-HT content. Neither lesion affected spinal (cervical-thoracic) 5-HT, nor NE in any of the brain areas assayed. It would appear that the dorsal and median raphe nuclei project 5-HT fibers into the forebrain but not to the spinal cord. The marger number of 5-HT fibers seems to originate in the dorsal raphe nucleus, which also seems to send a greater number of its 5-HT fibers to telecephalic than diencephalic structures. And, lastly, hippocampal and striatal 5-HT inputs apparently originate chiefly in the median and dorsal raphe nuclei, respectively.

The interpeduncular nuclear complex of selected rodents J

Article

Jan 1971

W R Ives

The internal differentiation of the interpeduncular nuclear complex was determined by quantitative cytoarchitectonic analysis in specimens from each of the suborders of Rodentia: rat (Rattus) of the Myomorpha; guinea pig (Cavia), chinchilla (Chinchilla), and capybara (Hadrochoerus) of the Caviamorpha; and the mountain beaver (Aplodonia) from the Sciuromorpha. In each of these rodents the constituent cells of the complex ae grouped into four spatially distinct subnuclei: pars lateralis (PL), pars medialis (PM), pars dorsalis magnocellularis (PDM), and pars dorsalis parvocellularis (PDP). There is a fifth subnucleus, the paramedian, in Chinchilla. Sample distributions, and means, of cell sixes were measured in each of these subdivisions in each species; and cell densities of each subdivision were determined in Rrattus. In all these animals PD had the smallest cells. PDM had the largest cells, except that in Hydrochoerus cells of PL, and in Cavia cells of PM, were as large as those of PDM. In Rattus and Aplodontia PL and PM were not different in cell size, and in Rattus their cells were of the same density. These findings cast doubt upon criteria previously used to subdivide this nuclear complex, and provide a consistent and replicable classification of its components.

Anatomical evidence for collateral branching of substantia nigra neurons: a combined horseradish peroxidase and [3H]wheat germ agglutinin axonal transport study in the rat

Article

Sep 1980
BRAIN RES

Ascending and cerebellar non-serotonergic projections from the nucleus raphe magnus of the rat

Article

Mar 1981
BRAIN RES

Non-serotonergic ascending and cerebellar projections of rat nucleus raphe magnus (RM) were disclosed using the horseradish peroxidase technique. The ascending bundle rose form rostral RM and was divided into two components; one ascended in raphe regions and the other in reticular formation. The former on innervated n. raphe dorsalis, etc., and the latter n. parafascicularis, zona incerta, etc. On the other hand, the axons innervating neocerebellum originated exclusively from caudal RM.

Connections of the corticomedial amygdala in the golden hamster. II. Efferents of the “olfactory amygdala”

Article

Mar 1981

The anterior cortical (C1) and posterolateral cortical (C2) nuclei of the amygdala are designated the “olfactory amygdala” because they each receive direct projections from the main olfactory bulb. The efferents of these nuclei were traced after sterotaxic placement of 1—5 μCi tritiated proline in the corticomedial amygdala of male golden hamsters. Following survival times of 12, 24, or 48 hours, 20 μm frozen sections of the brains were processed for light microscopic autoradiography. Efferents from C2 terminate in layers II and III of the olfactory tubercle and in layer lb of pars ventralis and pars medialis of the anterior olfactory nucleus. Fibers from this nucleus also project to layers I and II of the infralimbic cortex and to the molecular layer of the agranular insular cortex. More posteriorly, fibers from C2 teminate in layer I of the dorsolateral entorhinal cortex, and in the endopiriform nucleus. From C1, efferent fibers travel in the stria terminalis and terminate in the precommissural bed nucleus of the stria terminalis and in the mediobasal hypothalamus. Efferents from C1 also innervate the molecular layer of C2, the amygdalo-hippocampal area, and the adjacent piriform cortex. Neurons in both C1 and C2 project to the molecular layer of the medial amygdaloid nucleus and the posteromedial cortical nucleus of the amygdala, the plexiform layer of the ventral subiculum, and the molecular layer of the lateral entorhinal cortex.

Locus coerules neurons have axons that branch to the forebrain and cerebellum

Article

Dec 1981
BRAIN RES

Dennis Steindler

Utilizing a recently introduced double retrograde axonal tracing technique, the existence of collateralized axonal projections of locus coeruleus neurons to both forebrain and cerebellar cortical structures has been examined in adult mice. Following paired injection strategies using horseradish peroxidase and radiolabeled wheat germ agglutinin within portions of the forebrain and cerebellar cortices, substantial numbers of double labeled 'branched' neurons have been found that reside within all regions of the nucleus.

The nucleus raphe dorsalis of the rat and its projection upon the caudatoputamen. A combined cytoarchitectonic, immunohistochemical and retrograde transport study

Article

Feb 1981

The nucleus raphe dorsalis of the albino rat has been studied in the following three ways: (1) the cell mass was subjected to a detailed cytoarchitectonic analysis, based upon Nissl-stained material; (2) serotonin--as well as the noradrenaline--immunoreactive neurons present in the area of the nucleus raphe dorsalis were plotted; (3) following large injections of the fluorescent dye propidium iodide into the caudatoputamen complex, the cells in the nucleus raphe dorsalis projecting to this complex were labeled and subsequently stained with an antibody against serotonin. Cytoarchitectonic analysis showed that three cell types are present within the confines of the nucleus raphe dorsalis: small, medium and large. Moreover, differences in concentrations of cell bodies made it possible to subdivide the nucleus raphe dorsalis into four regions. Immunohistochemical analysis showed that the borders of the serotoninergic cell groups B6 and B7 of DAHLSTROM and FUXE do not coincide with those of the nucleus raphe dorsalis. Serotonin-immunoreactive perikarya in the nucleus raphe dorsalis were categorized as medium and large neurons; noradrenaline-immunoreactive neurons in the nucleus raphe dorsalis do all belong to the category--large neurons. With the combined use of immunofluorescence and fluorescent retrograde tracing, it was found that the projection from the nucleus raphe dorsalis to the caudatoputamen complex originates from serotoninergic as well as non-serotoninergic cells, both of which can be categorized as being medium-sized neurons. The data presented in this paper provides a guide for further studies of afferent and efferent connections of the nucleus raphe dorsalis and for electrophysiological experiments on its constituent neurons.

A Study of Overlap and Collateralization of Bulbar Reticular and Raphe Neurons which Project to the Spinal Cord and Diencephalon of the North American Opossum

Article

Feb 1984

Labeled neurons were found in most reticular and raphe nuclei of the opossum's medulla after horseradish peroxidase (HRP) injections into either the spinal cord or diencephalon. The labeling produced by spinal injections differed somewhat from that produced by diencephalic ones, but HRP-positive neurons were found in what appeared to be comparable areas. In order to study the intermingling of bulbospinal and bulbodiencephalic neurons more directly, we used fluorescent markers (true blue, nuclear yellow and diamidino yellow dihydrochloride) in double-labeling experiments. When injections of one dye into the spinal cord were combined with injections of another into the diencephalon, neurons labeled by one or the other marker were found to be intermingled in most reticular and raphe nuclei. The double-labeling experiments also showed that some neurons of the bulbar reticular formation and raphe project to both the spinal cord and diencephalon, presumably by axonal collaterals. The number of such neurons represents a somewhat larger proportion of those projecting to the diencephalon than to the spinal cord. It appears, however, that most neurons project either to the spinal cord or to the diencephalon and do not provide collaterals to both areas.

Electrophysiological Evidence of Concurrent Dorsal Raphe Input to Caudate, Septum, Habenula, Thalamus Hippocampus, Cerebellum and Olfactory Bulb

Article

Feb 1983

Field potentials evoked by electrical stimulation of the dorsal raphe nucleus (DR) were recorded in eight forebrain structures in the rat. The areas were chosen chiefly for their anatomical connections to the DR and included the olfactory bulb (OB), caudate nucleus (CN), lateral septum (Spt), lateral habenula (Hb), parafascicularis (PF), ventral thalamus (VT), hippocampus-CA1 (Hipp), and cerebellum (CB). DR stimulation evoked an initial biphasic positive-negative wave form at similar latencies in each of the eight structures. A later positive-negative-positive wave was evoked in only six structures: CN, Spt, Hb, PF, VT, and Hipp. The amplitudes and latencies of the peaks of the later wave forms varied among structures. A depth profile recording procedure created by moving the DR stimulating electrode showed that the responses in the remote structures occurred only when the stimulating electrode was located in the DR. Bursting the DR at 20 Hz for 5-20 min caused a decrease in all components of the response. The evoked potential amplitude returned to baseline levels 5-30 min after cessation of stimulation. The results indicates that the dorsal raphe has a concurrent input to many areas of the brain receiving 5HT afferents and that DR stimulation can modulate the neuronal activity in these regions.

Serotonergic and non-serotonergic projections from the nucleus raphe dorsalis to the caudate-putamen complex in the rat, studied by a combined immunofluorescence and fluorescent retrograde axonal labeling technique

Article

Oct 1980
NEUROSCI LETT

Immunofluorescence, induced in serotonergic neurons, was combined with retrograde axonal tracing by propidium iodide. Evidence is presented for the existence of serotonergic as well as non-serotonergic projections from the nucleus raphe dorsalis to the caudate-putamen complex.

The morphology of intracellularly labeled rat subthalamic neurons: A light microscopic analysis

Article

Apr 1983

Light microscopic analysis of rat subthalamic (STH) neurons which were intracellularly labeled with horseradish peroxidase, following the acquisition of electrophysiological data, revealed the following: (1) The somata of STH neurons were polygonal or oval with occasionally a few somatic spines. Usually three or four primary dendrites arose from the soma. Dendritic trunks tapered slightly and divided into long, thin, sparsely spined branches. Dendrites of some STH neurons extended into the cerebral peduncle. (2) Reconstruction of the dendritic field was made in three different planes. In either sagittal or frontal planes, the dendritic field was usually oval and the long axis was parallel to the main axis of STH. In the horizontal plane, the dendritic field of all neurons was polygonal. (3) The axons of all the neurons analyzed originated from the soma and were traced beyond the borders of STH, thus indicating that they were projection neurons. All the parent axons bifurcated at least once. After bifurcation, one axon branch coursed dorsolaterally within the cerebral peduncle and terminated in the globus pallidus. The other branch coursed caudally or mediocaudally and arborized in the substantia nigra. Frequently, the axon branches projecting toward the globus pallidus emitted fine axon collaterals within the entopeduncular nucleus. (4) About one-half of the analyzed STH neurons had intranuclear axon collaterals. The neurons with intranuclear collaterals had a higher dendritic tips/stems ratio than neurons without intranuclear collaterals. This observation indicated that STH neurons could be divided into two groups according to their axonal morphology. (5) The axonal terminal arborization observed in all the target sites (i.e., globus pallidus, entopeduncular nucleus, STH, and substantia nigra) were formed with varicose collateral branches which also gave rise to short filaments with beaded endings. Some of these projection neurons could therefore communicate with the target neurons in the globus pallidus, substantia nigra, entopeduncular nucleus, as well as STH through their collateral system.

Pallidal inputs to subthalamus: Intracellular analysis

Article

May 1983
BRAIN RES

Neuronal responses of the subthalamic nucleus (STH) to stimulation of the globus pallidus (GP) and the substantia nigra (SN) were studied by intracellular recording in the decorticated rat. (1) GP and SN stimulation evoked antidromic spikes in STH neurons with a mean latency of 1.2 ms and 1.1 ms, respectively. Based on the above latencies, the mean conduction velocity of the STH neurons projecting toward GP was estimated to be 2.5 m/s, and that toward SN was 1.4 m/s. Many STH neurons could be activated following stimulation of both GP and SN, indicating that single STH neurons project to two diversely distant areas. In spite of differences in conduction distance of GP and SN from STH, differences in the conduction velocities of bifurcating axons make it possible for a simultaneous arrival of impulses in the target areas to which these STH neurons project. (2) GP stimulation evoked short duration (5-24 ms) hyperpolarizing potentials which were usually followed by depolarizing potentials with durations of 10-20 ms. These potentials were tested by intracellular current applications and intracellular injections of chloride ions. The results indicated that the hyper- and depolarizing potentials were IPSPs and EPSPs respectively. These IPSPs were considered to be monosynaptic in nature since changes in the stimulus intensities of GP did not alter the latency of IPSPs. The mean latency of the IPSPs was 1.3 ms. Based on the above mean latency the mean conduction velocity of GP axons projecting to STH was estimated to be 3.8 m/s. (3) Analysis of electrical properties of STH neurons indicated that: (i) input resistance estimated by a current-voltage relationship ranged from 9 to 28 M omega; (ii) the membrane showed rectification in the hyperpolarizing direction; (iii) direct stimulation of neurons by depolarizing current pulses produced repetitive firings with frequencies up to 500 Hz. (4) Morphology of the recorded STH neurons was identified by intracellular labeling of neurons with horseradish peroxidase. Light microscopic analysis indicated that the recorded neurons were Golgi type I neurons with bifurcating axons projecting toward GP and SN.

Immunohistochemical evidence for the presence of γ-aminobutyric acid and serotonin in one nerve cell. A study on the raphe nuclei of the rat using antibodies to glutamate decarboxylase and serotonin

Article

Oct 1983
BRAIN RES

A specific and sensitive double immunocytochemical staining for the visualization of glutamate decarboxylase (GAD) and serotonin (5-HT) on the same brain section is developed. GAD is detected with specific GAD-antibodies by means of the unlabeled antibody enzyme, peroxidase anti-peroxidase, and serotonin with an antibody against the BSA-serotonin conjugate by an indirect immunofluorescent staining. The coexistence of GAD and 5-HT in the same perikaryon is demonstrated by a peroxidase reaction superimposed on fluorescent compounds. Cell bodies containing both antigens are observed in each raphe nuclei. However, the nucleus raphe dorsalis exhibits the largest number of cells containing either GAD alone or GAD and 5-HT together. An intracellular interaction between the metabolism of GABA and serotonin could be reasonably expected. The interactions between GABAergic and serotonergic systems must be thought of in terms of intracellular and/or transynaptic controls.

Combined immunocytochemistry and autoradiographic retrograde axonal tracing for identification of transmitters of projection neurons

Article

Dec 1983
J NEUROSCI METH

A double labeling method is described which combines immunocytochemistry for identification of the neurotransmitter serotonin with autoradiographic retrograde axonal tracing using wheat germ agglutinin (WGA), N-[acetyl-3H]. The permanence, sensitivity, and distinctness of the two labels provide a valuable means for analyzing transmitter-identified projection neurons in the central nervous system. Combined immunohistochemical/autoradiographic preparations, following injections of WGA, N-[acetyl-3H] in the caudate-putamen of mice, revealed large numbers of serotonergic and fewer non-serotonergic raphe-striatal projection neurons.

Rapid and simple determination of delivery after iontophoretic and pressure injections of radiolabeled tracer substances

Article

May 1983
J NEUROSCI METH

A fluorographic method is described using X-ray film analysis for the determination of delivery of radiolabeled tracer substances both in Agar plates and in tissue sections. This method is most useful in neuroanatomical autoradiographic studies for providing rapid identification of delivery, placement, and extent of an injection site after iontophoresis or pressure injections of radiolabeled axonal tracer substances.

N-[acetyl-3H] wheat germ agglutinin: Anatomical and biochemical studies of a sensitive bidirectionally transported axonal tracer

Article

Oct 1983
NEUROSCIENCE

Both retrograde and anterograde autoradiographic axonal labeling were associated with the appropriate pathways after injections of N-[acetyl-³H] wheat germ agglutinin within numerous structures in the central nervous system of mice. Sections processed for light microscopic autoradiography following the placement of injections mainly within the neocortex, neostriatum, or cerebellum have revealed patterns of bidirectional axonal labeling in various thalamic, monoaminergic, deep cerebellar and precerebellar nuclei that were similar to that seen after similarly placed horseradish peroxidase injections. Small injections of N-[acetyl-³H] wheat germ agglutinin, by way of an extremely limited extracellular spread of the tracer, yield large amounts of autoradiographic retrograde and anterograde axonal labeling. High specific activities, in addition to the sensitivity displayed by this tracer, allow the use of relatively short autoradiographic exposure times that still lead to an extensive signal over labeled neurons.

Differences in the labeling of axons of passage by wheat germ agglutinin after uptake by cut peripheral versus injections within CNS

Article

Nov 1982
BRAIN RES

Dennis Steindler

Injections of a radiolabeled derivative of wheat germ agglutinin (WGA) in the subcortical white matter of the cerebral cortex in mice do not give rise to autoradiographic labeling of axon systems coursing through this fiber mass. Exposing the cut-end of mouse tibial nerve to WGA does, however, produce labeling within dorsal root ganglia and the spinal cord. These findings are discussed with consideration for dissimilarity in mode of central versus peripheral administration of the tracer, as well as in the light of potential relative differences in the uptake and transport of WGA and HRP.

Differential uptake of peroxidase (HRP) and peroxidase-lectin (HRP-WGA) conjugate injected in the nodose ganglion of cat

Article

Oct 1982

A comparison was made of the uptake and consequent axonal transport of peroxidase and peroxidase-lectin conjugate injected in low concentrations (0.167%) in the nodose ganglion of cats. At the light microscopic level horseradish peroxidase (HRP)-wheat germ agglutinin (WGA) intensely labeled only central terminal fields of vagal afferents (anterograde), while free HRP only labeled perikarya in vagal motor nuclei (passing retrograde). Low concentrations of these proteins, in addition to normal diffusion equilibria, permit a differential distribution of those species demonstrating some affinity for cell membranes. We attribute these differences in the uptake of HRP and HRP-WGA to the selective affinity of WGA for cell surface receptors (n-acetyl glucosamine) on the plasma membrane. This results in a greater number of WGA molecules coupled to HRP being internalized in any given endocytotic event compared to free HRP. The fractionation of efferent and passing fiber populations within a nodosal injection site can be discriminated with these different preparations.

Identification of serotonin and non-serotonin-containing neurons of the mid-brain raphe projecting to the entorhinal area and the hippocampal formation. A combined immunohistochemical and fluorescent retrograde tracing study in the rat brain

Article

May 1982
NEUROSCIENCE

We have studied the localization of serotonin- and non-serotonin-containing cell bodies in the midbrain raphe nuclei that project to the entorhinal area and the hippocampal formation in the rat brain, using the technique of combined retrograde fluorescent tracing and immunohistochemistry on the same tissue section. The branching properties of these neurons were studied by retrograde double labelling using two fluorochromes which emit fluorescence with different spectral characteristics. After injections of granular blue or propidium iodide into the medial entorhinal area, retrogradely-labelled cells were found situated bilaterally in the caudal half of the dorsal raphe nucleus, the medial part of the median raphe and throughout the rostrocaudal extension of the nucleus reticularis tegmentipontis. Injections placed successively more laterally in the entorhinal area labelled progressively less cells contralaterally in the dorsal raphe and the reticular tegmental nucleus of the pons. After fluorochrome injections into the dorsal part of the hippocampal formation, retrogradely-labelled cells were found in the caudal part of the dorsal raphe, in the peripheral part of the median raphe and to a minor extent in the medial part of this nucleus, but not in the nucleus reticularis tegmentipontis. The experiments with double retrograde fluorescent tracing showed that the raphe nuclei do not send bilateral projections to the entorhinal area in spite of the fact that many of these cells are located contralateral to the injected hemisphere in single labelling experiments. Injections of the fluorochromes into the entorhinal area and hippocampal formation showed that at least 10% of the raphe cells project to both areas simultaneously. Analysis of sections incubated with antiserum to serotonin showed that a majority of the retrogradely-labelled versus serotonin-immunoreactive cells was found to vary within different parts of the individual raphe nuclei: the ventromedial part of the dorsal, the medial part of the median and the nucleus reticularis tegmentipontis being the highest. The findings indicate that both serotonin- and non-serotonin-containing neurons in the raphe innervate the hippocampal region, that these projections may be crossed but not bilateral, and that the same neuron in the raphe may influence the neural activity in the entorhinal area and the hippocampus simultaneously.

The organization of divergent axonal projections from the midbrain raphe nuclei in the rat

Abstract

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