[Progress in pineal research].
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Animals↗
Biomedical subjects
Y Shiotani
Publications and source records attributed to Y Shiotani.
Immunohistochemical localization of pituitary adenylate cyclase-activating polypeptide (PACAP) in the adrenal medulla of the rat.
Localization of PACAP in rat adrenal glands was examined by light and electron microscopic immunohistochemistry using a specific antiserum to PACAP 38, R0831. In the light microscopic study, PACAP immunoreactivity was observed in some cell groups in the medulla, but not in the cortex. In comparison with adjacent sections stained with antisera to catecholamine synthesizing enzymes, PACAP-positive cells were immunoreactive to tyrosine hydroxylase and dopamine beta-hydroxylase, but not to phenylethanolamine-N-methyltransferase, suggesting that they were coincident with noradrenaline secreting cells. In the electron microscopic study using the ABC method, DAB reaction products were diffusely distributed in the cytoplasmic matrix of PACAP-positive cells, without intense accumulation on the secretory granules. The splanchnic nerve terminals were PACAP negative. In postembedding immunohistochemistry, gold particles were localized diffusely in the cytoplasma, but not aggregated on the secretory granules. It was suggested that PACAP would localize in the cytoplasmic matrix of noradrenaline cells and stimulate the catecholamine synthesis and release in the adrenal medulla.
Adenylyl Cyclases↗
Localization of pituitary adenylate cyclase-activating polypeptide (PACAP) in the hypothalamus-pituitary system in rats: light and electron microscopic immunocytochemical studies.
The localization of pituitary adenylate cyclase-activating polypeptide (PACAP) in the hypothalamus-pituitary system in rats was examined in light and electron microscopic immunocytochemistry using a specific antiserum to synthetic PACAP 1-38 (R0831). In light microscopic study, intensely PACAP-immunostained perikarya were observed in the supraoptic and paraventricular magnocellular nucleus in the hypothalamus. In the median eminence, many immunoreactive nerve fibers were observed in the internal layer, but a few immunoreactive terminals were noticed in the external layer. In the pituitary gland, numerous immunoreactive nerve fibers were observed in the posterior lobe. In the intermediate lobe, moderately immunostained cells were observed, but in the anterior lobe no immunostained cells were noticed. In electron microscopic study, PACAP-immunoreactivity was examined by avidin-biotin peroxidase complex method. In the perikarya of the supraoptic and paraventricular magnocellular nucleus, DAB-reaction products were distributed diffusely in the cytoplasmic matrix, frequently attaching to the rough-surfaced endoplasmic reticulum. In the nerve terminals of the posterior lobe, reaction products were observed among the secretory granules, but sometimes upon them. In the cells of the intermediate lobe, reaction products were also distributed in the cytoplasmic matrix.
Animals↗
Involvement of specific placental antigen X-P2 in rat olfaction: an immunohistochemical study in the olfactory bulb.
Human placental antigen X-P2 (hPAX-P2), an antigen complex associated with cytochrome P-450 of aromatase within estrogen synthesizing tissues, has been reported to be present in a distinct group of rat primary olfactory receptors involved in suckling behavior. In this study, most of the mitral and tufted cells in the rat olfactory bulb were found to possess hPAX-P2 immunoreactivity. This suggests that the activity of these cells can be hormonally modulated and that hPAX-P2 is involved in rat olfaction not only at the receptor level but also at integrative brain levels via the secondary projecting neurons of the olfactory pathway.
Animals↗
Effects of constant light and darkness on the intrapineal neurons of golden hamsters, stained for tyrosine hydroxylase. A morphometric analysis.
Tyrosine hydroxylase (TH)-positive neurons (TH neurons) were found in the pineal gland of golden hamsters. To examine possible relations between TH neurons and environmental light, we kept male animals under constant light (LL) and darkness (DD) for a week, and morphometrically compared the number, size, and immunoreactivity of TH neurons with those of control animals kept under 12L/12D (LD), using an image processor, Nexus 6400. In LL animals, the number of TH neurons/mm2 of pineal tissue and each cell area were decreased, and immunoreactivity to TH was less than in LD animals. In DD animals, the number of TH neurons and each cell area were increased, and immunoreactivity decreased slightly. These data suggested that environmental light affected the TH neurons, and the amount of TH in the neurons would be decreased by LL, but increased by DD.
Animals↗
Screening of aromatase-containing neurons in rat forebrain: an immunohistochemical study with antibody against human placental antigen X-P2 (hPAX-P2).
Aromatase-containing neurons were immunohistochemically examined in rat brains by using a polyclonal antibody against human placental antigen. The antibody recognizes cytochrome P-450 portion of aromatase, an enzyme converting androgen to estrogen. A large group of strongly immunoreactive cells was identified in the ventral pallidum, which extends caudally from the area surrounding the islands of Calleja. Other strongly or moderately stained cell groups were observed in the cerebral cortex, the amygdaloid area, the nucleus of the diagonal band, and the area anterior to the posterior commissure. Only a few stained cells were present in the medial preoptic region. These findings cast doubt upon the previous assumption, based on biochemical analysis of tissue samples, that the center of the aromatizing system is in the medial preoptic region. They indicate instead that most aromatase-containing neurons of rats lie within the ventral pallidum ventromedially adjacent to the preoptic area.
Animals↗
Interaction between cholinergic neurons and substance P or calcitonin gene-related peptide terminals of the rat sacral intermediolateral nucleus: double immunostaining at the light and electron microscopic levels.
The relationships both between cholinergic neurons and substance P (SP) and between cholinergic neurons and calcitonin gene-related peptide (CGRP) terminals were examined in the rat sacral intermediolateral nucleus at the light and electron microscopic levels by means of double-immunostaining methods. Cholinergic neurons were labeled by a monoclonal antibody to choline acetyltransferase (CAT) with the avidin-biotin technique and stained bluish-green by indolyl-beta-galactoside reaction products with beta-galactosidase as a marker. On the same sections, SP or CGRP fibers were labeled by polyclonal antisera to SP or CGRP after application of the peroxidase-antiperoxidase (PAP) method and stained brown by the p-dimethylaminoazobenzene (DAB) reaction. After embedding in Epon, light and electron microscopic sections were examined. At the light microscopic level, CGRP-like immunoreactive (CGRP-I) fibers and SP-like immunoreactive (SP-I) fibers were found to pass through the lateral edge of the dorsal horn and then into the dorsal region of the sacral intermediolateral nucleus. In addition, SP-I fibers also extend from the dorsolateral funiculus into the entire sacral intermediolateral region. At the electron microscopic level, many axosomatic and axodendritic synapses were found between CAT-I structures and SP-I terminals in the intermediolateral nucleus, whereas most of the CGRP-I terminals in this area made axodendritic synapses with CAT-I dendrites. These results indicate that cholinergic neurons in the sacral intermediolateral nucleus receive direct synaptic input from SP-I and CGRP-I terminals.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
"Necklace olfactory glomeruli" form unique components of the rat primary olfactory system.
A distinct subset of rat primary olfactory neurons was identified immunohistochemically by means of a polyclonal antibody against human placental antigen X-P2 (hPAX-P2), an incompletely characterized substance found in all estrogen-biosynthetic organs. The subset of olfactory receptor cells was distributed widely over the olfactory epithelium with some degree of concentration on the dorsocaudal walls of nasal subcavities. The subset formed unique "necklace olfactory glomeruli," which were composed of seven to nine solitary glomeruli located in the caudal end of the olfactory bulb. One of them was located in the "modified glomerular complex" reported to be involved in rat suckling behavior. The projectional patterns of the necklace olfactory system, albeit diffuse, indicated some degree of spatial correspondence between zones of olfactory epithelium and specific glomeruli. Axons emanating from neighboring cells can project to several glomerular loci. From the necklace olfactory system, an average of 150-200 receptor cells were estimated to converge onto a single necklace glomerulus.
Animals↗
Ontogeny of tyrosine hydroxylase-positive but dopamine beta-hydroxylase-negative neuron-like cells in the pineal gland of golden hamsters.
The ontogeny of tyrosine hydroxylase (TH)-positive but dopamine beta-hydroxylase (DBH)-negative neuron-like cells in the pineal gland of golden hamsters was investigated by double immunostaining. These cells first appeared on the 6th postnatal day and thereafter increased in number. On the other hand, TH- and DBH-positive nerve fibers, probably originating from the superior cervical ganglion, were already present in the pineal gland at birth.
Animals↗
Enkephalin fibers synapse on cholinergic neurons in the rat sacral intermediolateral nucleus: a double-immunostaining at the light and electron microscopic levels.
Relationships between leucine-enkephalin fibers and cholinergic neurons in the rat sacral intermediolateral nucleus were examined by light and electron microscopy using double-immunostaining method. Cholinergic neurons in the sacral intermediolateral nucleus were labeled by a rat-mouse monoclonal antibody to choline acetyltransferase and stained bluish green with 5-bromo-4-chloro-3-indolyl-beta-D- galactoside reaction products using beta-galactosidase as a marker. On the same sections, leucine-enkephalin fibers were labeled by a rabbit polyclonal antiserum to leucine-enkephalin and stained brown by diaminobenzidine reaction products using peroxidase as a marker. After embedding in Epon, the sections were examined in light and electron microscopes. In the light microscope, choline acetyltransferase-like immunoreactive cells were seen in the sacral intermediolateral nucleus. In the same region, leucine-enkephalin-like immunoreactive cells. In the electron microscope, 5-bromo-4-chloro-3-indolyl-beta-D-galactoside reaction products were in the form of coarse electron dense deposits in the choline acetyltransferase-like immunoreactive structures and could be distinguished from the much finer grained diaminobenzidine reaction products. Choline acetyltransferase-like immunoreactive neurons received synaptic inputs from leucine-enkephalin fibers-like immunoreactive terminals. These findings suggest that leucine-enkephalin fibers may affect the activity of cholinergic parasympathetic preganglionic neurons.
Animals↗
Immunohistochemical demonstration of tyrosine hydroxylase (TH)-positive but dopamine beta-hydroxylase (DBH)-negative neuron-like cells in the pineal gland of golden hamsters.
Double immunostaining for tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) in the pineal gland revealed many TH-positive but DBH-negative, neuron-like cells in golden hamsters, but not in rats and gerbils. They were scattered in the meshwork of TH- and DBH-positive nerve fibers throughout the parenchyma. Their cell bodies were oval or spindle-shaped (maximum diameter: 15-20 micron). They had sometimes one or two short cell processes. Bilateral superior cervical ganglionectomy diminished strikingly the number of TH- and DBH-positive nerve fibers, but did not affect the number of TH-positive cells.
Animals↗
Direct adrenergic inputs to sacral autonomic neurons: using a double-immunostaining method at the light and electron microscopic levels.
Relationships between adrenergic fibers and cholinergic neurons were examined in the rat sacral intermediolateral nucleus using a double-immunostaining method at light and electron microscopic levels. Adrenergic fibers were immunohistochemically stained brown by peroxidase reaction, and cholinergic neurons in the same sections were stained bluish green by beta-galactosidase reaction. In the light microscope, many cholinergic neurons were seen in the intermediolateral nucleus and some of them were surrounded by adrenergic fibers. In the electron microscope, adrenergic fibers made synapses with the cholinergic neurons in the intermediolateral nucleus. These results suggest that adrenergic fibers directly influence the activity of the sacral parasympathetic preganglionic neurons.
Animals↗
Analysis of the rat interpeduncular subnuclei by immunocytochemical double-staining for enkephalin and substance P, with some reference to the coexistence of both peptides.
The rat interpeduncular nucleus (IPN) was immunocytochemically double-stained for enkephalin (ENK) and substance P (SP) on the same sections. On the basis of both peptidergic distribution patterns and topographic relationship, the IPN was divided into nine subnuclei and one cap: the rostral subnucleus (IP-R), the central subnucleus (IP-C), the rostral-lateral subnucleus (IP-RL), the main lateral subnucleus (IP-L), the caudal-lateral subnucleus (IP-CL), the dorsal-lateral subnucleus (IP-DL), the dorsal-medial subnucleus (IP-DM), the apical subnucleus (IP-A), the intermediate subnucleus (IP-I), and the dorsal cap (IP-Cap). As the descriptions of the IP-RL, IP-L, and IP-CL were inconsistent with previous reports, they were reevaluated; the IP-RL was proposed as the region situated in the lateral portion at rostral levels and characterized by the lack of ENK and SP immunoreactive structures, the IP-L as the region situated throughout the rostrocaudal extent in the lateral portion of the IPN and containing the highest density of SP immunoreactive fibers but no ENK immunoreactive fibers, and the IP-CL as the region situated just laterocaudal to the IP-L in the caudal pole of the IPN and containing ENK immunoreactive cells and fibers but no SP immunoreactive structures. Our results also showed that some cells in the IP-R have both ENK and SP immunoreactivity. This coexistence was observed in some small spherical cells of the IP-R, but rarely in larger oval-shaped cells, which occasionally showed only ENK immunoreactivity. In addition, paired ENK immunoreactive fiber bundles entering the IP-R were found to run just rostral to the paired SP immunoreactive columns, both of which composed parts of the interpedunculotegmental tract. A three-dimensional model representing the subnuclear organization of the IPN was proposed on the basis of the present results.
Brain Mapping↗
Histaminergic nerve fibers in the median eminence and hypophysis of rats demonstrated immunocytochemically with antibodies against histidine decarboxylase and histamine.
Histaminergic fibers in the median eminence and hypophysis of rats were examined immunocytochemically with antibodies against histidine decarboxylase (HDC), the sole histamine-synthesizing enzyme, and against histamine itself. A similar distribution of immunoreactive fibers was observed with these two antibodies. In the median eminence, immunoreactive fibers were mainly located in the internal layer and could be traced to the posterior lobe of the hypophysis. A few fibers were detected in the external layer of the median eminence, but none in the anterior or intermediate lobe of the hypophysis. These observations suggest that neuronal histamine may take part in regulation of the hypothalamo-neurohypophysial neuroendocrine system in rats.
Animals↗
Interaction between adrenergic fibers and intermediate cholinergic neurons in the rat spinal cord: a new double-immunostaining method for correlated light and electron microscopic observations.
Relationships between cholinergic neurons and adrenergic fibers in the intermediate region of the rat thoracic spinal cord were examined using a new immunohistochemical double-staining method for light and electron microscopic observations. Cholinergic neurons were labeled by a monoclonal antibody to choline acetyltransferase and stained bluish green by 5-bromo-4-chloro-3-indolyl-beta-D-galactoside reaction products using beta-galactosidase as a marker. On the same sections, adrenergic fibers were labeled by a polyclonal antiserum to phenyl-ethanolamine-N-methyltransferase and stained brown by diaminobenzidine reaction products using peroxidase as a marker. After embedding in Epon, the sections were examined in the light and electron microscopes. In the light microscope, choline acetyltransferase-like immunoreactive cells were seen in the four discrete areas of the intermediate region: the principal intermediolateral nucleus, the central autonomic nucleus, the intercalated nucleus and the funicular intermediolateral nucleus. These cell groups seemed to be connected to each other by their processes, and they showed a "ladder-like appearance" as a whole. Phenylethanolamine-N-methyltransferase-like immunoreactive fibers were present only along this "ladder-like structure" and were the most rich in the principal intermediolateral nucleus. In the electron microscope, some of the choline acetyltransferase-like immunoreactive neurons, which were identified by light micrographs, were found to receive synaptic inputs from phenylethanolamine-N-methyltransferase-like immunoreactive boutons in the principal intermediolateral nucleus. These findings suggest that the adrenergic axons in the principal intermediolateral nucleus directly affect the activity of the cholinergic preganglionic sympathetic neurons.
Adrenergic Fibers↗
Neurotensin-containing projections from the retrosplenial cortex to the anterior ventral thalamic nucleus in the rat.
The neurotensin-containing projections from the retrosplenial cortex to the anterior ventral thalamus were demonstrated by electrolytic lesion studies and fluorescent retrograde tracing combined with immunocytochemistry. Three-to-five-day-old rats were used, because the immunoreactivity of neurotensin fibers in anterior ventral thalamus was the highest at this age. When neurotensin-containing neurons located in layer VI of the retrosplenial cortex were unilaterally destroyed by applying an electrolytic current to the retrosplenial area, the neurotensin fibers in the ipsilateral anterior ventral thalamus decreased dramatically. Unilateral injection of a fluorescent retrograde tracer, Fast Blue, into the anterior ventral thalamus, labeled neurons in the ipsilateral retrosplenial cortex, and many of these cells also had neurotensin-like immunoreactivity. These results suggested that a major origin of the neurotensin fibers in the anterior ventral thalamus was in the ipsilateral retrosplenial granular cortex.
Amidines↗
Coexistence of substance P- and enkephalin-like peptides in single neurons of the rat hypothalamus.
The distribution of substance P- and enkephalin-like immunoreactivity in single cells were examined by the double immunofluorescence method. Substance P- and leucine-enkephalin-like compounds coexisted within individual neurons of some hypothalamic areas such as the medial preoptic area, anterior hypothalamic area, perifornical area, lateral hypothalamic area, premammillary nuclei and posterior hypothalamic nucleus, although they did not coexist in the majority of immunoreactive cells.
Animals↗
The histaminergic innervation of the mesencephalic nucleus of the trigeminal nerve in rat brain: a light and electron microscopical study.
Histaminergic fibers in the mesencephalic nucleus of the trigeminal nerve of Long-Evans rats were examined by light and electron microscopy after peroxidase-antiperoxidase immunocytochemical staining for histidine decarboxylase (HDC) as a marker. By light microscopy, neurons in the mesencephalic nucleus of the trigeminal nerve were seen to be surrounded by a number of HDC-like immunoreactive (HDCI) fibers, suggesting the presence of axo-somatic contact. This finding was supported by immunoelectron microscopic demonstration of synaptic contact of some HDCI fibers with the soma of the neurons in this nucleus. These findings indicate that histamine is involved in the sensory regulation of movement of the masticatory muscles at the level of the trigeminal mesencephalic nucleus.
Animals↗