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PubMed1988

Colocalization of GABA and glycine immunoreactivities in a subset of retinal neurons in tiger salamander.

Alternate serial 1 micron Durcupan resin sections of tiger salamander retina were stained with antisera against GABA and glycine using postembed immunocytochemical techniques. Although the vast majority of neurons were labeled by either GABA or glycine antiserum, a small percentage of presumed amacrine cells in the inner nuclear layer and cells in the ganglion cell layer were clearly labeled by both antisera, indicative of colocalization of endogenous GABA and glycine. Although there is a greater than 90% chance that a labeled cell will be clearly labeled for either GABA or glycine immunoreactivity, the possibility for cotransmission of two inhibitory transmitters must be considered for a small percentage of these retinal neurons.

Animals↗

PubMed1988

Colocalization of GABA and [Met]enkephalin-Arg6-Gly7-Leu8 in the rat cerebellum.

The distribution of [Met]enkephalin-Arg6-Gly7-Leu8-like immunoreactivity (MEAGL-LI) in the rat cerebellum was investigated by peroxidase anti-peroxidase immunocytochemistry using specific antiserum against MEAGL. MEAGL-LI positive neuronal perikarya were distributed in the granular layer, and they seemed to correspond to Golgi cells from their size and location. In addition, diffusely and weakly stained neuronal perikarya were also observed in the molecular layer. Immunoreactive fibers and terminals were found in the granular layer. Furthermore, examination of serial frozen sections (4-6 micron in thickness) from rats pretreated with colchicine clarified the colocalization of gamma-aminobutyric acid (GABA) and MEAGL in Golgi cells but not in the stellate cells.

Animals↗

PubMed1988

Distribution and colocalization of delta sleep inducing peptide (DSIP) with corticotropin-like intermediate lobe peptide (CLIP) in the human hypophysis.

DSIP and CLIP [ACTH(18-39)] immunoreactive (IR) neurons and fibers were examined in the human hypophysis and pituitary stalk using immunmohistofluorescence and peroxidase-antiperoxidase methods. Double-stained and adjacent stained sections demonstrate that DSIP is colocalized in about 75% of CLIP-IR-like cells in the anterior pituitary and in residual intermediate lobe cells. Only few CLIP-IR-like fibers are observed in the posterior lobe. On the contrary, a high density of DSIP-IR fibers is visualized in the stalk. It is suggested that DSIP acts as a sleep promoting factor (one of many other actions) and that CLIP increases the paradoxical sleep, so that these two peptides could play a role in the regulating system of the sleep-waking cycle.

Adrenocorticotropic Hormone↗

PubMed1990

Colocalization of calbindin and GABA in medial nucleus of the trapezoid body of the rat.

Using immunocytochemical methods, both calbindin and GABA were found to be colocalized in the somas of all the cells of the medial nucleus of the trapezoid body (NMTB) of the rat auditory system. In the lateral superior olive (LSO), calbindin was also found in the terminals but not in the cells. Some terminal labelling was found in the medial superior olive (MSO). GABA was also found in the somas of some cells in both LSO and MSO, but most of the labelling was in terminals. In the rat, calbindin appears to be more involved in a pathway that detects interaural intensity differences.

Animals↗

PubMed1992

Colocalization of nitric oxide synthase and NADPH-diaphorase in the myenteric plexus of the rat gut.

The pattern of distribution and colocalization of nitric oxide-synthase (NOS) and NADPH-diaphorase in the myenteric plexus of whole-mount preparations of the antrum, duodenum, ileum, caecum, proximal colon and distal colon of the rat were investigated using immunohistochemical and histochemical staining techniques. Almost all the myenteric neurons that were NOS-positive in all regions of the gut examined were also stained for NADPH-diaphorase. However, in the stomach, duodenum and ileum, only a few of the NOS-positive nerve fibres in the tertiary and secondary plexuses and circular muscle layer were also stained for NADPH-diaphorase, whereas in the caecum and distal colon almost all the NOS-positive nerve fibres were also stained for NADPH-diaphorase. The results in the present study are consistent with the view that nitric oxide (NO) has a mediating role in gastrointestinal neurotransmission.

Amino Acid Oxidoreductases↗

PubMed1992

Effects of CNS stimulants on the in vivo release of the colocalized transmitters, dopamine and neurotensin, from rat prefrontal cortex.

The effect of CNS stimulant drugs on the in vivo release of the colocalized neurotransmitters dopamine and neurotensin in rat prefrontal cortex was studied using microdialysis. Amphetamine, methylphenidate and nomifensine all increased extracellular fluid (ECF) levels of dopamine; however, their effects of neurotensin varied. Amphetamine increased both ECF dopamine (514 +/- 82% of basal) and neurotensin (350 +/- 49% of basal); however, the neurotensin increase lagged behind the increase in dopamine suggesting a possible trans-synaptic effect. Methylphenidate increased both dopamine and neurotensin (226 +/- 26% and 151 +/- 14% of basal respectively) co-synchronously, suggesting exocytosis of vesicles containing both dopamine and neurotensin. The nomifensine-induced increase in dopamine (202 +/- 23% of basal) was similar to that of methylphenidate, whereas the increase in neurotensin was significantly delayed and of lower magnitude (134 +/- 20% of basal). These data suggest that dopamine and neurotensin in part share a common releaseable pool in the prefrontal cortex. Moreover, dopamine may act presynaptically to increase neurotensin release and the different behavioral profiles of these psychostimulants may in part relate to their different effects on neurotensin release.

Analysis of Variance↗

PubMed1993

Neurons in rat cerebral cortex that synthesize nitric oxide: NADPH diaphorase histochemistry, NOS immunocytochemistry, and colocalization with GABA.

Neurons that stain for NADPH diaphorase, which colocalizes with nitric oxide synthase (NOS), are scattered uniformly across neocortex, and denser in entorhinal cortex. In the primary sensorimotor cortex, 0.5-2% of neurons contain NOS. These are most numerous in layers II-III, whereas NOS-positive fibers are concentrated in layers IV and VI. Most stained neurons are aspiny bipolar cells. Some in deep layers are multipolar; very few are pyramidal-shaped. In layer IV, NOS-positive neurons and their dendrites are confined to the septa between barrels. Retrograde tracing experiments demonstrate that NOS-positive cells are local circuit neurons. Double staining demonstrates that NOS-positive neurons also contain GABA.

Amino Acid Oxidoreductases↗

PubMed1993

Colocalization of GABA and glycine in the rabbit oculomotor nucleus.

In the present study we examined the possible colocalization of the inhibitory neurotransmitters glycine and GABA in the oculomotor nucleus of the rabbit. Serial sections were processed alternately for glycine and GABA postembedding immuno-cytochemistry. Ultrastructural analysis revealed that all terminals that showed glycine-positive immunoreactivity were also GABA positive; up to 5% of the GABA-positive terminals were also glycine positive.

Animals↗

PubMed1994

Colocalization of binding sites for somatostatin, muscarine and nicotine on cultured neurones of rat neocortex, cerebellum, brain stem and spinal cord: combined autoradiographic and immunohistochemical studies.

The cellular localization of binding sites for [125I]1-tyramine somatostatin ([125I]SS) was studied in explant cultures of rat CNS by autoradiography. In cultures from cortex, brain stem and spinal cord many neurones revealed binding sites for the peptide whereas in cerebellar cultures only little binding of [125I]SS was observed. In addition to neurones, astrocytes were also labelled by the peptide. By combined immunohistochemical and autoradiographic techniques, it was demonstrated that the majority of neurones which expressed binding sites for [125I]SS were also immunostained by the monoclonal cholinergic muscarinic or nicotinic receptor antibodies (M 35 and W 6, respectively), providing evidence for a colocalization of cholinergic and somatostatin receptors on the neuronal membrane.

Animals↗

PubMed1989

Immunohistochemical colocalization of delta sleep-inducing peptide and luteinizing hormone-releasing hormone in rabbit brain neurons.

The anatomical distributions of luteinizing hormone-releasing hormone and delta sleep-inducing peptide immunoreactivity in the rabbit brain were studied by indirect immunofluorescence technique. The comparison of adjacent serial sections, one being immunolabeled with an antiserum to luteinizing hormone-releasing hormone, the other with an antiserum to delta sleep-inducing peptide, showed that the respective distribution patterns of immunoreactivity exhibited a remarkable overlap through the basal forebrain and hypothalamic regions. A sequential double-immunolabelling (elution-restaining method) clearly indicated that all the luteinizing hormone-releasing hormone-immunoreactive cell bodies displayed delta sleep-inducing peptide immunoreactivity. These cell bodies were sparse and mainly located throughout the septal-preoptico-suprachiasmatic region and the ventrolateral hypothalamus. The colocalization of luteinizing hormone-releasing hormone and delta sleep-inducing peptide immunoreactivity was also observed in many fibres supplying all these brain regions and terminal areas such as the organum vasculosum of the lamina terminalis, the subfornical organ, the median eminence and the pituitary stalk. These neuroanatomical findings are suggestive of interaction between delta sleep-inducing peptide and luteinizing hormone-releasing hormone in various brain areas including some circumventricular organs.

Brain↗

PubMed1986

Neuronal colocalization of adenosine deaminase, monoamine oxidase, galanin and 5-hydroxytryptophan uptake in the tuberomammillary nucleus of the rat.

Neurons of the tuberomammillary nucleus (TM) in the rat have previously been shown to contain the enzymes adenosine deaminase (ADA), histidine decarboxylase (HDC) and glutamate decarboxylase (GAD). Some neurons coextensive with this cell group also exhibit immunoreactivity for the neuropeptide galanin, express monoamine oxidase activity (MAO), or display the ability to accumulate and decarboxylate 5-hydroxytryptophan (5-HTP). Histochemical and immunohistochemical techniques were used to determine the extent to which these neurochemical properties are colocalized in neurons immunoreactive for adenosine deaminase. Galanin was found to coexist with ADA in about 45% of the neurons in the TM. In addition, a large number of cells immunoreactive for galanin alone were observed in the posterior hypothalamus outside the confines of TM. Neurons displaying MAO activity formed a subpopulation of those immunoreactive for ADA; all neurons containing MAO also contained ADA whereas only 60% of the ADA-immunoreactive cells were reactive for MAO. Approximately 20% of ADA-immunoreactive neurons represented nearly all cells having 5-HTP uptake capability. However, a very few cells in TM showing 5-HTP uptake capability appeared to be devoid of ADA immunoreactivity. These results demonstrate that although neurons of TM are homogeneous with respect to a number of possible neurotransmitters markers and associated enzymes, these neurons are heterogeneous with respect to their expression of galanin, MAO and 5-HTP uptake. In certain respects the segregation of histochemical properties within TM correlates with previous histochemical work by others, and suggests the possibility of functional diversity of TM.

5-Hydroxytryptophan↗

PubMed1996

Activation of anterior lobe corticotrophs by electroacupuncture or noxious stimulation in the anaesthetized rat, as shown by colocalization of Fos protein with ACTH and beta-endorphin and increased hormone release.

A marked expression of the c-fos proto-oncogene has been recently reported in cells of the anterior lobe of the pituitary gland in rats subject to electroacupuncture or noxious thermal stimulation under pentobarbital anaesthesia. The present study was undertaken to identify the activated pituitary cells. Following both kinds of stimulation, most Fos-immunoreactive anterior lobe cells showed colocalization with adrenocorticotropic hormone or beta-endorphin immunoreactivity. No c-fos expression occurred in pituitary cells immunoreactive for growth hormone, prolactin, luteinizing hormone, or thyrotropin-stimulating hormone. A marked rise of adrenocorticotropic hormone and beta-endorphin concentrations occurred in plasma. In the hypothalamus, c-fos expression was increased in the mediobasal nuclei-namely, the arcuate nucleus-and in the paraventricular nucleus, but more in the former. It is suggested that somatosensory noxious input, or the partly noxious input evoked by electroacupuncture, activate the hypothalamo-pituitary-adrenocortical axis as in common forms of stress, but with a specific activation of the mediobasal hypothalamic nuclei and no stimulation of intermediate lobe cells. Opiate release from the pituitary gland may contribute to acupuncture analgesia or the intrinsic antinociceptive reactions triggered by noxious stimulation.

Adrenocorticotropic Hormone↗

PubMed1991

Neurochemical hypothesis: participation by aluminum in producing critical mass of colocalized errors in brain leads to neurological disease.

1. Aluminum is an established neurotoxin. Prolonged exposure to even low levels of aluminum permit its chelation and subsequent transport to brain where it is non-uniformly distributed. 2. Available evidence suggests that (i) aluminum interferes with glucose metabolism by inhibiting hexokinase and glucose-6-phosphate dehydrogenase; (ii) it binds to calmodulin and affects numerous phosphorylation-dephosphorylation reactions; (iii) it binds to transferrin and ferritin, affects the function of these proteins which in turn affect iron metabolism. 3. Thus accumulation of aluminum-induced metabolic errors colocalized in specific areas of the brain may lead to neurological disorders.

Aluminum↗

PubMed1991

Colocalization of immunoreactive oxytocin, vasopressin and interleukin-1 in human thymic epithelial neuroendocrine cells.

Monoclonal antibodies to oxytocin (OT) and vasopressin (VP) revealed some positively staining stromal cells in the subcapsular cortex and in the medulla of the human thymus. We further demonstrated that these cells are a subset of epithelial endocrine cells and also contain immunoreactive interleukin-1 together with the neuropeptides. In addition, the thymic cells stained by monoclonal antibodies directed to the cyclic part of oxytocin or vasopressin also contained some immunoreactive neurophysins. These data support the concept of intrathymic synthesis of neurohypophyseal-like peptides fitting the hypothalamic model. However, we observed that, contrary to the situation in the brain, OT- and VP-like peptides colocalized in the same thymic cells. Furthermore, one monoclonal antibody, specific for the tail part of oxytocin, did not label thymic cells. Therefore, thymic neuropeptide(s) could be related to, but distinct from, authentic OT and VP. These observations suggest some molecular differences between hypothalamic and thymic oxytocin biosynthetic pathways which need to be further investigated.

Antibodies, Monoclonal↗

PubMed1994

Somatostatin, galanin and peptide histidine isoleucine in the newborn and adult human trigeminal ganglion and spinal nucleus: immunohistochemistry, neuronal morphometry and colocalization with substance P.

By means of indirect immunofluorescence the neuropeptides somatostatin, galanin and peptide histidine isoleucine were localized in cell bodies, nerve fibres and terminal-like elements in the ganglion and spinal nucleus of the human trigeminal nerve in perinatal and adult ages. No immunoreactivity to vasoactive intestinal polypeptide was observed. In the gasserian ganglion somatostatin-, galanin- and peptide histidine isoleucine-containing neurons and nerve fibres occurred frequently in pre- and full-term newborns, but were scarce to absent in adults. Somatostatin- and galanin-positive pericellular basket-like structures around non-immunoreactive perikarya were observed in newborn specimens. Immunoreactivity to somatostatin, galanin and peptide histidine isoleucine labelled nerve fibers and punctate and felt-like nerve terminals in the pars interpolaris and subnucleus caudalis of the spinal trigeminal nucleus, with immunostaining and distribution patterns characteristic for each peptide. In addition, somatostatin-containing neuronal cell bodies frequently were detected. At variance with those containing somatostatin, the number of galanin- and peptide histidine isoleucine-like immunoreactive elements were dramatically reduced in the adult tissue compared to the newborn one. Double immunostaining revealed that each of the three peptides partially colocalizes with substance P, the degree of coexistence being very low for somatostatin/substance P and high for galanin/substance P and peptide histidine isoleucine/substance P both in the gasserian ganglion and in the spinal nucleus. The results obtained suggest that somatostatin, galanin and peptide histidine isoleucine may play functional roles in primary sensory neurons and at the first synaptic level of the human trigeminal sensory system.

Aged↗

PubMed1994

Partial colocalization of the GABAA receptor with parvalbumin and calbindin D-28K in neurons of the visual cortex and the dorsal lateral geniculate nucleus of the cat.

Monoclonal antibodies to a synthetic peptide fragment of the beta 1-subunit of the bovine central GABAA/benzodiazepine receptor were used to investigate immunocytochemically the distribution of this receptor in the visual system of the cat. Labeled neurons were observed in all layers of the visual cortex and the dorsal lateral geniculate nucleus. About half of the total cortical or geniculate neuronal population was found to be positive. To further identify immunocytochemically these GABAA receptor expressing cells, double stainings were undertaken with, on one hand, the monoclonal antibodies directed against the receptor complex, and on the other hand polyclonal antisera directed against cat muscle parvalbumin or chicken calbindin D-28K. A high degree of colocalization between either of the two calcium binding proteins and the GABAA receptor was found in the upper layers (I, II and III) of the visual cortex and in the A and C laminae of the dorsal lateral geniculate nucleus; all calbindin D-28K-positive cells were immunoreactive for the GABAA receptor. The parvalbumin-positive cells, scattered throughout all layers of the dorsal lateral geniculate nucleus and the visual cortex, except cortical layer I, were also all positive for the GABAA receptor. However, a large proportion of all GABAA receptor bearing cells were negative for one of the calcium binding proteins.

Animals↗

PubMed1996

Immunohistochemical mapping of nitric oxide synthase in the rat hypothalamus and colocalization with neuropeptides.

The localization and distribution of nitric oxide synthase in the hypothalamus have been studied with an immunohistochemical technique using antibodies to neuronal rat nitric oxide synthase. Subsequent double-labeling experiments examined the colocalization patterns of nitric oxide synthase and several peptides. Our results demonstrate a widespread occurrence of nitric oxide synthase-immunoreactive nerve cell bodies and processes throughout the hypothalamus, especially in various parts of the preoptic region, in the supraoptic and paraventricular nuclei, the lateral hypothalamic area, the ventromedial and dorsomedial nuclei, the arcuate nucleus and various parts of the mammillary region. Double labeling experiments showed that nitric oxide synthase-like immunoreactivity coexists with substance P-like immunoreactivity in the medial preoptic area, with oxytocin-, cholecystokinin-and galanin message-associated peptide-like immunoreactivity in the supraoptic nucleus, with enkephalin, oxytocin- and corticotropin releasing factor-like immunoreactivity in the paraventricular nucleus and with enkephalin-like immunoreactivity in the arcuate nucleus. Furthermore, in the ventromedial nucleus, nitric oxide synthase-like immunoreactivity coexisted with enkephalin-, substance P-, and somatostatin-like immunoreactivity, and in the dorsomedial nucleus with enkephalin-, galanin message-associated peptide-and substance P-like immunoreactivity. In the mammillary region nitric oxide synthase-like immunoreactivity coexisted with enkephalin-, cholecystokinin-, and substance P-like immunoreactivity. Among these neuropeptides, enkephalin and substance P were most frequently found in nitric oxide synthase-immunoreactive neurons. We conclude that nitric oxide synthase-immunoreactive neurons contain neuropeptides in various parts of the hypothalamus, and that nitric oxide in the hypothalamus may be involved in a variety of neuroendocrine and autonomic functions.

Animals↗

PubMed1996

Colocalization of NOS and SOD1 in neurofilament accumulation within motor neurons of amyotrophic lateral sclerosis: an immunohistochemical study.

Peroxynitrite, formed from nitric oxide and superoxide, may affect neurofilament assembly and cause neurofilament accumulation in motoneurons. This hypothesis may reconcile the mutations of two genes: superoxide dismutase-1 in some patients with familial amyotrophic lateral sclerosis, and the gene for the heavy neurofilament in some patients with sporadic amyotrophic lateral sclerosis previously reported. We found colocalization of superoxide dismutase-1 and nitric oxide synthase in the foci of neurofilament accumulation as 'conglomerates' in upper motor neurons and 'axonal spheroids' in lower motor neurons. In addition, all the specific molecules related to the reactions, including calmodulin, 3', 5'-cyclic guanosine-monophosphate, citrulline, and nitrotyrosine were found strongly immunopositive in the site of neurofilament accumulation. Our data support the view that the neurofilament aggregates are tightly linked with superoxide dismutase-1 and nitric oxide synthase activities. Both enzymes may focally contribute to peroxynitrite formation at light neurofilament, which is rich in both tyrosine and arginine residues and hence considered as the vulnerable site for nitrotyrosine formation. Nitrotyrosine is known to inhibit phosphorylation and if it impairs phosphorylation of neurofilament subunits, either light or heavy, may alter the slow axonal transport culminating in proximo-distal accumulation of NF and slowly progressive motoneuron death.

Amyotrophic Lateral Sclerosis↗