Fig 5 - uploaded by Vassil Papantchev
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Electron microscope features of the nNOS-ir structures of the neurophil from different major and accessory vestibular nuclei. (a) Longitudinal section of the NOS I-immunopositive dendrite. Note the association of the immunoproduct with the dendritic neurotubules. An axo-spinous synaptic contact is also present. SB—synaptic bouton, asterisk—dendritic spine. Preparation form LVN. (b) Cross-section through out the nNOS-ir dendrite filled with immunoproduct. Note the great number of unlabeled synaptic terminals (asterisks). Preparation form SVN. (c) nNOS-ir large round (LR) terminal bouton, forming an axodendritic synapse with labeled dendrite (D). Preparation form MVN. (d) nNOS-ir large round (LR) terminal bouton, forming an axo-somatic synapse with a labeled neuron. Preparation form SVN. (e) Different in size and shape nNOS-ir myelinated axons (mAX). Preparation form IVN. In all parts of the figure, bar = 0.25 lm
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Nitric oxide is a unique neurotransmitter, which participates in many physiological and pathological processes in the organism. Nevertheless, there are little data about the neuronal nitric oxide synthase immunoreactivity (nNOS-ir) in the vestibular complex of a cat. In this respect, the aims of this study were to: (1) demonstrate nNOS-ir in the ne...
Contexts in source publication
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... immunoproduct was seen in dendrites, myelinated axons, and terminal boutons throughout the nuclei of VNC (Fig. ...
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... dendrites were different in size-from small (0.5-1 lm) to large (up to 3 lm). The immunoproduct was associated with the axial neurofilaments in the dendrites (Fig. 5a). A great number of nNOS-ir SB terminated on the labeled dendrites, together with many unlabeled ones (Fig. 5a, ...
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... dendrites were different in size-from small (0.5-1 lm) to large (up to 3 lm). The immunoproduct was associated with the axial neurofilaments in the dendrites (Fig. 5a). A great number of nNOS-ir SB terminated on the labeled dendrites, together with many unlabeled ones (Fig. 5a, ...
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... nNOS-ir SB were usually large, containing a great number of round vesicles and many mitochondria (Fig. 5c, d). This synaptic structure is typical for large round SB. Usually the large round SB performed a great number of asymmetric synaptic contacts (Fig. 5c, d). Rarely axo-spinous synapses were present (Fig. ...
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... nNOS-ir SB were usually large, containing a great number of round vesicles and many mitochondria (Fig. 5c, d). This synaptic structure is typical for large round SB. Usually the large round SB performed a great number of asymmetric synaptic contacts (Fig. 5c, d). Rarely axo-spinous synapses were present (Fig. ...
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... nNOS-ir SB were usually large, containing a great number of round vesicles and many mitochondria (Fig. 5c, d). This synaptic structure is typical for large round SB. Usually the large round SB performed a great number of asymmetric synaptic contacts (Fig. 5c, d). Rarely axo-spinous synapses were present (Fig. ...
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... immunoproduct was observed in myelinated axons (Fig. 5e). Labeled axo-somatic synaptic contacts were also found (Fig. ...
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Citations
... Nitric oxide (NO) has been recognized as a unique gaseous neurotransmitter (Garthwaite, 1991;Holstein et al., 2001;Martinelli et al., 2002), and neurons expressing the NOsynthesizing enzyme, nitric oxide synthase (NOS), exist in certain cell populations of the brain (Snyder, 1992;Olmos et al., 2005). Nitric oxide takes part in many important physiologic processes such as synaptic plasticity in the hippocampus (Krushkov et al., 1996), as well as eye movements (Moreno-López et al., 1996, blood pressure control (Maeda et al., 1999), and the functional integrity of the claustrum (Hinova-Palova et al., 1997), thalamus (Krushkov et al., 1996), inferior colliculus (Paloff and Hinova-Palova, 1998), and vestibular complex (Papantchev et al., 2005(Papantchev et al., , 2006. Histochemical mapping studies of the claustrum reveal NOS and NADPH-d activity (Vincent and Kimura, 1992;Paloff et al., 1994;Rodrigo et al., 1994;Hinova-Palova et al., 1997, 2013Paloff and Hinova-Palova, 1998;Paxinos and Watson, 1998;Vincent, 2000;Edelstein et al., 2012a,b). ...
... Within the last 30 years, several studies have demonstrated the presence of NO in the nervous system of a variety of vertebrates. Relative to morphological investigations, a number of immunocytochemical and histochemical studies have utilized antibodies against NOS or have used the NADPH-diaphorase histochemical technique (Mizukawa et al., 1989(Mizukawa et al., , 1990Dawson et al., 1991Dawson et al., , 1993Dawson et al., , 1994Hope et al., 1991;Rodrigo et al., 1994;Kullo et al., 1997;Soares et al., 2003;Marino and Cudeiro, 2003;Papantchev et al., 2005Papantchev et al., , 2006Sears et al., 2004;Seyidova et al., 2004). ...
... It is well known fact that neurons differing in shape and size may possess different functional attributes (Papantchev et al., 2005(Papantchev et al., , 2006. The primay focus of the present study was the comparative assessment of the topographical distribution, morphology and arborizations of NOS-ir neurons in the human and rat claustrum. ...
We compared the distribution, density and morphological characteristics of nitric oxide synthase-immunoreactive (NOS-ir) neurons in the rat and human claustrum. These neurons were categorized by diameter into three main types: large, medium and small. In the human claustrum, large neurons ranged from 26 to 40 μm in diameter, medium neurons from 20 to 25 μm and small neurons from 13 to 19 μm. In the rat claustrum, large neurons ranged from 19 to 23 μm in diameter, medium neurons from 15 to 18 μm and small neurons from 10 to 14 μm. The cell bodies of large and medium neurons varied broadly in shape – multipolar, elliptical, bipolar and irregular, consistent with a projection neuron phenotype. The small neurons were most seen as being oval or elliptical in shape, resembling an interneuron phenotype. Based on a quantitative comparison of their dendritic characteristics, the NOS-ir neurons of humans and rats displayed a statistically significant difference.
... Nitric oxide is readily identified using NADPH-d histochemistry and standard immunocytochemical procedures. NADPH-d-positive neurons and fibers are evident in many parts of the CNS across a variety of species, including humans (Mizukawa et al. 1989;Mizukawa 1990;Vincent and Kimura 1992;Moreno-Lopez et al. 1998;Saxon and Beitz 2000;Lysakowski and Singer 2000;Holstein et al. 2001;Martinelli et al. 2002;Papantchev et al. 2005Papantchev et al. , 2006Dzhambazova et al. 2008Dzhambazova et al. , 2011aLandzhov et al. 2011a, b;Landzhov and Dzhambazova 2012;Hinova-Palova et al. 2014;Edelstein et al. 2012a, b). Although there is an abundance of information on the presence of NADPH-d in the brain, data concerning its distribution within the IC and the detailed characterization of such are scant. ...
Using the nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) reaction with nitroblue tetrazolium, we provided a detailed investigation of the distribution, dimensional characteristics and morphology of NADPH-d-positive neurons in the three main subdivisions of the human inferior colliculus (IC): central nucleus, pericentral nucleus, and external nucleus. In accordance with their perikaryal diameter, dendritic and axonal morphology, these neurons were categorized as large (averaging up to 45 μm in diameter), medium (20-30 µm), small (13-16 µm) and very small (7-10 µm). Their morphological differences could contribute to varying functionality and processing capacity. Our results support the hypothesis that large and medium NADPH-d-positive cells represent projection neurons, while the small cells correspond to interneurons. Heretofore, the very small NADPH-d-positive neurons have not been described in any species. Their functions - and if they are indeed the smallest neurons in the IC of humans - remain to be clarified. Owing to their location, we posit that they are interneurons that connect the large NADPH-d-positive neurons and thereby serve as an anatomical substrate for information exchange and processing before feeding forward to higher brain centers. Our results also suggest that the broad distribution of nitric oxide (NO) synthesis in the human IC is closely tied to the neuromodulatory action of NO on collicular neurotransmitters such as GABA and glutamate, and to calcium-binding proteins such as parvalbumin. A deeper understanding of the relationship between NADPH-d-positive fibers in all IC connections and their colocalization with other neurotransmitters and calcium-binding proteins will assist in better defining the function of NO in the context of its interplay with the cerebral cortex, the sequelae of the aging process and neurodegenerative disorders.
... It has been established that neurons of differing shapes and sizes can have different functions (Papantchev et al., 2006). One of the aims of the present study was to identify whether NPY immunoproduct was present in possibly functionally distinct subpopulations of claustral neurons. ...
The claustrum is a telencephalic nucleus located ventrolateral to the basal ganglia in the mammalian brain. It has extensive reciprocal connectivity with most if not all of the cerebral cortex, in particular, primary sensory areas. However, despite renewed and growing interest amongst investigators, there remains a paucity of data concerning its peptidergic profile. The aim of the present study was to examine the presence, morphology, distribution and ultrastructure of neuropeptide Y-immunoreactive (NPY-ir) neurons and fibers in the claustrum of the cat. Ten adult healthy cats from both sexes were used. All animals received humane and ethical treatment in accordance with the Principles of Laboratory Animal Care. Subjects were irreversibly anesthetized and transcardially perfused with fixative solution containing glutaraldehyde and paraformaldehyde. Brains were promptly removed, postfixed and sectioned. Slices were incubated with polyclonal anti-NPY antibodies according to the standard Avidin-Biotin-Peroxidase Complex method adopted by our Department of Anatomy, Histology and Embryology. NPY-ir neurons and fibers were found to be diffusely distributed throughout the claustrum, with no obvious topographic or functional patterning other than larger numbers in its central/broadest part (stereotaxic planes A12-A16). Neurons were generally classified by diameter into three sizes: small (under 17μm), medium (17-25μm) and large (over 25μm). Staining density varied, with some neurons appearing darker than others. At the electron-microscopic level NPY immunoproduct was observed within neurons, dendrites and terminal boutons, each differing relative to their ultrastructural attributes. Two types of NPY-ir synaptic boutons were found. Lastly, it is of interest to note that gender-specific differences were not observed.
... Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPHd)-positive neurons and fibers are present in many parts of the nervous system in a variety of species, including humans (Mizukawa et al., 1989;Mizukawa, 1990;Vincent and Kimura, 1992;Druga and Syka, 1993;Valtschanoff et al., 1993;Yan et al., 1996;Hinova-Palova et al., 1997;Paloff and Hinova-Palova, 1998;Moreno-Lopez et al., 1998;Saxon and Beitz, 2000;Lysakowski and Singer, 2000;Holstein et al., 2001;Martinelli et al., 2002;Papantchev et al., 2005Papantchev et al., , 2006Edelstein et al., 2012a,b). Histochemical mapping studies of the mammalian brain reveal either neuronal nitric oxide synthase (NOS) or NADPHd activity in the claustrum (Vincent and Kimura, 1992;Paloff et al., 1994;Rodrigo et al., 1994;Switka et al., 1994;Hinova-Palova et al., 1997, 2013Paloff and Hinova-Palova, 1998;Paxinos and Watson, 1998;Vincent, 2000;Edelstein et al., 2012a,b). ...
We studied the topographical distribution and morphological characteristics of NADPH-diaphorase-positive neurons and fibers in the human claustrum. These neurons were seen to be heterogeneously distributed throughout the claustrum. Taking into account the size and shape of stained perikarya as well as dendritic and axonal characteristics, Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPHd)-positive neurons were categorized by diameter into three types: large, medium and small. Large neurons ranged from 25 to 35 μm in diameter and typically displayed elliptical or multipolar cell bodies. Medium neurons ranged from 20 to 25 μm in diameter and displayed multipolar, bipolar and irregular cell bodies. Small neurons ranged from 14 to 20 μm in diameter and most often displayed oval or elliptical cell bodies. Based on dendritic characteristics, these neurons were divided into spiny and aspiny subtypes. Our findings reveal two populations of NADPHd-positive neurons in the human claustrum-one comprised of large and medium cells consistent with a projection neuron phenotype, the other represented by small cells resembling the interneuron phenotype as defined by previous Golgi impregnation studies.
... A further series of rinses were performed according to the protocol used in the Department of Anatomy and Histology (Papantchev et al. 2006;Hinova-Palova et al. 2007;Papantchev 2008). The first rinse was in PBS, followed by Tris buffer (pH 7.6), which preceded the visualization of peroxidase activity using H 2 O 2 and 3,3 0 -diaminobenzidine as substrates. ...
... The procedure continued with three 2-min rinses in Tris buffer, followed by another three 2-min rinses in PBS. Sections for electron microscopy were postfixed with 1 % OsO 4 in PBS for 1 h, dehydrated in graded series of ethanol, and flat-embedded in Durcupan (Fluka, Buchs, Switzerland) between acetate sheets (Papantchev et al. 2006;Hinova-Palova et al. 2007;Papantchev 2008;Papantchev et al. 2009a, b). Sections were trimmed-out under a dissecting microscope and glued to epoxy blanks. ...
... Sections were trimmed-out under a dissecting microscope and glued to epoxy blanks. Thin sections were cut with an ultramicrotome (LKB, Stockholm-Bromma, Sweden), counterstained with uranyl acetate and lead citrate according to the protocol used in the Department of Anatomy and Histology (Papantchev et al. 2006;Hinova-Palova et al. 2007;Papantchev 2008;Papantchev et al. 2009a, b), and examined with an Hitachi H-500 transmission electron microscope (Hitachi, Tokyo, Japan). ...
The claustrum is a complex telencephalic structure owing to its reciprocal connectivity with most-if not all-cortical areas. However, there is a paucity of data in the literature concerning its histochemical components, including opioid peptide neurotransmitters. The aim of the present study was to examine the morphology, distribution and ultrastructure of leucine-enkephalin-immunoreactive (Leu-enk-ir) neurons and fibers in the dorsal claustrum (DC) of the cat. Seven healthy, adult male and female cats were used in our study. All animals received humane care. They were irreversibly anesthetized and transcardially perfused with fixative. Brains were removed, postfixed, blocked and sectioned. Sections were incubated with polyclonal anti-Leu-enk antibodies using the Avidin-Biotin-Peroxidase Complex method. Leu-enk-ir neurons and fibers were distributed throughout the DC. Some of the neurons were lightly-stained, while others were darkly-stained. Light-microscopically, they varied in shape: oval, fusiform, multipolar and irregular. With regard to size, they were categorized as small (15 μm or less in diameter), medium (16-20 μm in diameter) and large (21 μm or more in diameter). No specific pattern of regional distribution was found. On the electron microscope level, immunoproduct was observed in neurons, dendrites and terminal boutons. Different types of Leu-enk-ir neurons differ in their ultrastructural features, including two types of synaptic boutons. No gender-specific features were observed. In conclusion, it is our hope that our study will serve to contribute to a better understanding of the functional neuroanatomy of the DC in the cat, and that it can be extrapolated and applied to other mammals, including humans.
... Nitric oxide (NO) is a unique neurotransmitter, which participates in many physiologi cal and pathological processes in the organism. It was synthesized by the family o f enzymes called nitric oxide synthase (NOS), which belong to three subtypes -neuronal (nNOS), inducible (iNOS) and endothelial (eN O S )) and had widespread distribution in the brain [6,9,10,11]. ...
Nitric oxide (NO) synthesized by the enzyme nitric oxide synthase (NOS) affects the secretion o f stress hormones and NO system is a stress-limiting system. Also, NO is involved in NO-molecular ways, which affect through auto regulation different signaling molecules - like opioids, endocannabinoids and others. Many stress models have been reported to affect the levels of nitric oxide and stimulate the expression of NOS engaged in their synthesis. Stimulation of opioid receptors within the periaqueductal gray (PAG) activates descending opioid and noradrenaline inhibitory pathways and suppresses nociception.
Because PAG has been identified as region that mediates the response to different stressful paradigms and contains distinct, longitudinally organised neural substrates, our goal was to investigate the effect o f 3 hours immobilization stress (IS) on NO activity in rat ventrolateral PAG (vlPAG) by a histochemical procedure for nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d). According to obtained data NADPH-d reactive neurons in rat’s vlPAG and NO activity respectively was increased by stress model mentioned above.
Key words: Nitric oxide, immobilization stress, vlPAG
... Nitric oxide (NO) is a unique gasous neurotransmitter (Holstein et al. 2001, Martinelli et al. 2002. It was shown that it takes part in many processes in the central nervous system like synaptic plasticity in the hippocampus (Krushkov et al. 1996), claustral functioning (Hinova-Palova et al. 1997), eye movements (Moreno-Lopez et al. 1996, thalamic functioning (Krushkov et al. 1996), blood pressure control (Maeda et al. 1999), inferior colliculus functioning , vestibular complex functioning (Papantchev et al., , 2006 and so forth. NO is synthesized by the enzyme Nitric oxide synthase (NOS). ...
... During the last years a lot of studies on NOS and NO have been performed (Mizukawa et al. 1989, Mizukawa 1990, Dawson et al. 1991a, 1991b, 1994, Hope et al. 1991, Rodrigo et al. 1994, Kullo et al. 1997, Soares et al. 2003, Marino and Cudeiro 2003, Papantchev et al. 2003, 2006, Sears et al. 2004, Seyidova et al. 2004. Nevertheless there are little data about the neuronal Nitric oxide synthase immunoreactive (nNOS-ir) neurons and fibers in the dorsal claustrum (DC) of a cat. ...
... All animals received humane care in compliance with the ''Principles of laboratory animal care'' formulated by the National Society for Medical Research and the ''Guide for the care and use of laboratory animals'' prepared by the National Institute of Health (NIH publication No. 86-23, revised 1996). Our perfusion protocol was described in details elsewhere (Papantchev et al. 2006). In brief-all animals were deeply anesthetized with intraperitoneal injection of Urethan (40 mg/kg) and transcardialy perfused with 500 ml heparinized saline followed by 3,000 ml phosphate buffered saline (PBS; pH 7.4) containing 2.5% glutaraldehyde and 4% parafomaldehyde. ...
Nitric oxide is a unique neurotransmitter, which participates in many physiological and pathological processes in the organism. Nevertheless there are little data about the neuronal Nitric Oxide Synthase immunoreactive (nNOS-ir) neurons and fibers in the dorsal claustrum (DC) of a cat. In this respect the aims of this study were: (1) to demonstrate nNOS-ir in the neurons and fibers of the DC; (2) to describe their light microscopic morphology and distribution; (3) to investigate and analyze the ultrastructure of the nNOS-ir neurons, fibers and synaptic terminals; (4) to verify whether the nNOS-ir neurons consist a specific subpopulation of claustral neurons; (5) to verify whether the nNOS-ir neurons have a specific pattern of organization throughout the DC. For demonstration of the nNOS-ir the Avidin-Biotin-Peroxidase Complex method was applied. Immunopositive for nNOS neurons and fibers were present in all parts of DC. On the light microscope level nNOS-ir neurons were different in shape and size. According to the latter they were divided into three groups-small (with diameter under 15 microm), medium-sized (with diameter from 16 to 20 microm) and large (with diameter over 21 microm). Some of nNOS-ir neurons were lightly-stained while others were darkly-stained. On the electron microscope level the immunoproduct was observed in neurons, dendrites and terminal boutons. Different types of nNOS-ir neurons differ according to their ultrastructural features. Three types of nNOS-ir synaptic boutons were found. As a conclusion we hope that the present study will contribute to a better understanding of the functioning of the DC in cat and that some of the data presented could be extrapolated to other mammals, including human.
Stress is known to exert an influence on neuroendocrine, autonomic, hormonal, and immune functioning. Various stress models have been reported to induce analgesia. This is a phenomenon referred to as stress-induced analgesia (SIA). One of the mechanisms known to play a part in the response of an organism to stress is activation of the endogenous opioid system. Endogenous opioid peptides take part in various functions as hormones or neuromodulators. Tyr-W-MIF-1 and Tyr-K-MIF-1 are neuropeptides, neuromodulators which are able to inhibit the expression of some forms of SIA. Nitric oxide (NO) plays an important role in initiation and maintenance of pain. It is also known that acute and chronic stress induce biochemical changes affecting both pain threshold and behaviour. Thus, endogenous opioid peptides and nitric oxide (NO) mediated a wide variety of physiological processes including pain transmission and SIA. The aim of the study was to investigate the effects of Tyr-W-MIF-1 and Tyr-K-MIF-1 on nociception. after immobilization, hot and cold stress (IS, CS and HS) and NO-involvement. L-Arginine, L-NAME and SIN-1 were used. Nociception was measured in male Wistar rats by paw pressure (PP) test. In conclusion we suggest that there is a different kind of involvement of endogenous nitric oxide in the mechanisms of nociception of Tyr-W-MIF-1 and Tyr-K-MIF-1 after immobilization, cold and hot stress.
The vestibular nuclear complex (VNC) is an intricate structure subdivided into major and accessory nuclei all with different functions. In general the studies of the normal ultrastructure of the VNC are few in number. Furthermore, most data are contradicting and there are a lot of "white spots" in the entire scientifi{dotless}c literature concerning this issue. The aim of the present study was to examine the normal ultrastructure of the VNC of the cat. Eight adult healthy cats from both sexes were used. All experiments were performed in compliance with the current local laws. All animals received humane care. They were anaesthetized and transcardially perfused with fi{dotless}xative solution. Brains were removed, postfi{dotless}xed and cut. Slices were prepared for standard electron microscopy according to the standard prescription of the Department of Anatomy and Histology. According to their ultrastructural characteristics the neurons were subdivided into three groups - small, medium-sized and large. The ultrastructural features show that small neurons are interneurons, while large neurons are projection neurons. During the present study we observed all neuronal types in all nuclei. We were not able to demonstrate any sex-related differences. Our results show that different nuclei have relatively simple and uniform ultrastructure. On the basis of our observations and accumulated literary data we can conclude that the ultrastructure alone could not explain functional heterogeneity of the complex.
Carbon monoxide (CO) and nitric oxide (NO) are two endogenously produced gases that can function as second messenger molecules in the nervous system. The enzyme systems responsible for CO and NO biosynthesis are heme oxygenase (HO) and nitric oxide synthase (NOS), respectively. The present study was undertaken to examine the distribution of HO-2 and NOS of the trigeminal primary afferent neurons of the rat, located in the trigeminal ganglion (TG) and mesencephalic trigeminal nucleus (MTN), using histochemistry and immunohistochemistry. NADPH-d staining was found in most neurons in TG. The intensely NADPH-d-stained neurons were small- or medium-sized, while the large-sized neurons were less intensely stained. Immunocytochemistry for HO-2 revealed that almost all neurons in TG expressed HO-2, but they did not appear cell size-specific pattern. NADPH-d and HO-2 positive neurons appeared the same pattern, which was NADPH-d activity and HO-2 expression progressively declined from the caudal to rostral part of the MTN. A double staining revealed that the colocalization of NADPH-d/HO-2 neurons was 97.3% in TG and 97.6% in MTN. The remarkable parallels between NADPH-d and HO-2 suggest that NO and CO are likely neurotransmitters and mediate the orofacial nociception and sensory feedback of the masticatory reflex arc together.
