Sadao Shiosaka

Distribution of the histaminergic neuron system in the central nervous system of rats; a fluorescent immunohistochemical analysis with histidine decar☐ylase as a marker

The distribution of histidine decarboxylase-like immunoreactivity (HDCI) in the rat central nervous system was studied by the indirect immunofluorescence technique. HDCI cell bodies were concentrated in the posterior hypothalamic area, such as in the tuberal magnocellular nucleus, caudal magnocellular nucleus, posterior hypothalamic nucleus and lateral hypothalamus just lateral to the fasciculus mammillothalamicus at the level of the posterior hypothalamic nucleus. Extensive networks of HDCI fibers of various densities were found in many areas of the brain; they were particularly dense in the hypothalamus but were also found in the following areas: rostrally in the cerebral cortex, olfactory nuclei, medial amygdaloid nucleus, n. tractus diagonalis, and bed nucleus of the stria terminalis, and caudally in the central gray matter of the midbrain and pons, auditory system, n. vestibularis medialis, n. originis nervi facialis, n. parabrachialis, n. commissuralis, n. tractus solitarii, and n. raphe dorsalis.

Coexistence of calcitonin gene-related peptide and substance P-like peptide in single cells of the trigeminal ganglion of the rat: immunohistochemical analysis

The localization of calcitonin gene-related peptide (CGRP) and substance P (SP) in the rat trigeminal ganglion was examined by means of the indirect immunofluorescent method. About 40% of neurons in the ganglion contained CGRP-like immunoreactivity (CGRPI), while about 20% of neurons showed SP-like immunoreactivity (SPI). In serial sections, nearly all the SPI neurons contained CGRPI.

Topographic localization of calcitonin gene-related peptide in the rat brain: An immunohistochemical analysis

Abstract The distribution of immunoreactive calcitonin gene-related peptide in the rat brain was investigated by means of an indirect immunofluorescence method. In addition to previously reported calcitonin gene-related peptide-like immunoreactive structure-containing sites such as the nucleus ambiguus, nucleus originis nervi facialis, nucleus originis nervi hypoglossi, nucleus peripeduncularis and nucleus parabrachialis, the present study demonstrated a far wider distribution of calcitonin gene-related peptide-like immunoreactive structure-containing cells in the rat brain, i.e. the nucleus hypothalamicus lateralis. nucleus ventromedialis thalami, colliculus superior, lemniscus lateralis, gyrus dentatus. nucleus olivaris superior, nucleus tractus solitarii, nucleus cuneiformis, nucleus parabigeminalis and a proportion of the Purkinje cells. We have also demonstrated a more extensive network of calcitonin gene-related peptide-like immunoreactive fibers distributed in various areas throughout the rat brain than has been reported previously such as the colliculus inferior, nucleus olivaris superior, nucleus vestibularis lateralis and inferions, and nucleus cochlearis dorsalis and ventralis, etc.

Ontogeny of substance P-containing neuron system of the rat: Immunohistochemical analysis—I. Forebrain and upper brain stem

The ontogeny of substance P-containing neuron system in the forebrain and upper brain stem of the rat was investigated by means of the indirect immunofluorescence technique. Substance P-positive structures first appeared in the primordium of the epithalamus and the area which surrounded the commissura posterior of the rat fetus corresponding to gestational day 14 (10-12 mm embryos). On and after gestational day 14, substance P-positive structures gradually made their appearance in various areas of the forebrain and upper brain stem. Substance P-positive structures thus continued to increase in number and in density during the fetus and perinatal stage and showed histochemically maximum content at the stage between postnatal days 5 and 15. After then, substance P-positive neurons tended to decrease in number as the rats grew, while substance P-positive fibers maintained in general their strong immunoreactivity even in the adult rats. The present study demonstrates that substance P-positive structures appear at a very early ontogenetical stage. This suggests that substance P might play an important role in the development of the forebrain and upper brain stem in addition to its neurotransmitter or neuromodulator functions.

Immunohistochemical evidence for the coexistence of calcitonin gene-related peptide- and choline acetyltransferase-like immunoreactivity in neurons of the rat hypoglossal, facial and ambiguus nuclei

The present immunocytochemical study demonstrates that calcitonin gene-related peptide-like immunoreactivity (CGRPI) coexists with acetylcholine in single cells of hypoglossal, facial and ambiguus nuclei. The experiments were done using alternate frozen sections from relevant regions of the rat brain. We further show that CGRPI is localized in the nerve terminals that form neuromuscular junctions in the tongue muscles.

Evidence for the presence of a histaminergic neuron system in the rat brain: An immunohistochemical analysis

Histamine-containing cells in rats were identified by indirect immunofluorescent histochemistry using an antibody raised against histidine decarboxylase (HDC), the enzyme forming histamine, which was purified from fetal rat liver. HDC-like immunoreactive (HDCI) structures could be detected in the brain as well as in peritoneal mast cells and basal-granulated cells in deep crypts of the gastric mucosa of rats. Numerous HDCI neurons were found in the posterior hypothalamic area and HDCI nerve fibers with a varicose appearance of fluorescence were widely distributed in various regions of the brain.

Increase of basic fibroblast growth factor immunoreactivity and its mRNA level in rat brain following transient forebrain ischemia

SummaryWe examined the time course of basic fibroblast growth factor (bFGF) immunoreactivity and its mRNA level mainly in the hippocampus after transient forebrain ischemia using immunohistochemistry, enzyme immunoassay (EIA), Western blot analysis and in situ hybridization. Neuronal death in the hippocampal CA1 subfield was observed 72 h after 20 min of ischemia. The number of bFGF-immunoreactive(IR) cells increased 48 h–5 days after ischemia in all hippocampal regions. At 10 and 30 days, the bFGF-IR cells in the CA1 subfield had further increased in numbers and altered their morphology, enlarging and turning into typical reactive astrocytes with the advancing neuronal death in that area. In contrast, the number of bFGF-IR cells in other hippocampal regions had decreased 30 days after ischemia. The EIA study showed a drastic increase in bFGF levels in the hippocampus 48 h after ischemia (150% of that in normal rat) which was followed by further increases. In Western blot analysis, three immunoreactive bands whose molecular weights correspond to 18, 22 and 24 kDa were observed in normal rat and ischemia increased all their immunoreactivities. In the in situ hybridization study of the hippocampus, bFGF mRNA positive cells were observed in the CA1 subfield in which many bFGF-IR cells existed after ischemia. These data demonstrate that transient forebrain ischemia leads to an early and strong induction of bFGF synthesis in astrocytes, suggesting that the role of bFGF is related to the function of the reactive astrocytes which appear following brain injury.

Ascending components of the medial forebrain bundle from the lower brain stem in the rat, with special reference to raphe and catecholamine cell groups. A study by the HRP method

The afferent connection of the medial forebrain bundle (MFB) arising from the lower brain stem have been investigated by means of horseradish peroxidase (HRP) with sensitive substrate, the injection was made iontophoretically into MFB at various levels. After injection of HRP into MFB, a significant number of HRP-labeled neurons were observed in the following structures of the lower brain stem: (1) raphe nuclear group, (2) locus coeruleus, (3) n. laterodorsalis tegmenti, (4) parabrachial area, (5) A1, A2, A4, A5 and A7 areas where noradrenaline-containing neurons were disseminated, (6) A8, A9 and A10 areas which contain dopamine neurons, (7) surrounding area of the fasciculus longitudinalis medialis at the level of the n. propositus hypoglossi, (8) n. prepositus hypoglossi and (9) mesencephalic gray matter. As a rule, the ascending projections are ipsilateral and course in the medial part of MFB. Regarding the raphe nuclei, we have demonstrated that the caudal raphe nuclei, such as n. raphe magnus and obscurus (but not n. raphe pallidus), also send their axons to the hypothalamus. Particularly, the axons of n. raphe magnus ascend in MFB to reach the level of the preoptic or anterior septal areas. Furthermore, in accordance with previous reports, HRP-labeled cells were also identified in the n. raphe dorsalis, centralis superior and pontis, respectively. It should be further noted that labeled cells appeared in the n. linearis caudalis. In addition, the present study indicates a number of non-aminergic cell groups as sources of ascending mfb fibers. On the whole, the present study further clarified the organization of the components of the MFB ascending from the lower brain stem, and provided some additional anatomical substrates for the physiology of the control of the forebrain by the lower brain stem neurons.

Ontogeny of substance P-containing neuron system of the rat: Immunohistochemical analysis—II. Lower brain stem

Ontogeny of the substance P neuron system in the lower brain stem of the rat was investigated by means of the indirect immunofluorescence technique. Substance P-positive structures (cells and fibers) first appeared in the primordium of the ventral part of the nucleus tractus spinalis nervi trigemini (nVs), in the reticular formation between nucleus reticularis lateralis and nVs, and in the reticular formation between nVs and nucleus originis nervi facialis, respectively, at gestational day 14. After that time, substance P-positive structures made their appearance gradually in various areas of the lower brain stem. The present study demonstrates that substance P-positive structures appear at a very early ontogenetical stage, which suggests that substance P may play a role in the development of the lower brain stem in addition to its role as a neurotransmitter or neuromodulator.

Experimental immunohistochemical studies on the amygdalofugal peptidergic (substance P and somatostatin) fibers in the stria terminals of the rat

The amygdalofugal substance P (SP) and somatostatin (SRIF) neuron systems in the stria terminalis (ST) were investigated by means of the indirect immunofluorescence technique of Coons. SP- and SRIF-positive cells were mainly located in the area (Amc) between the central (ac) and medial (am) amygdaloid nuclei. Some extended medially into the am and laterally into the ac. Destruction of the Amc resulted in a marked reduction of SP- and SRIF-positive fibers in the ST. Furthermore, a substantial decrease in SP-positive fibers was seen in the dorsal part of the bed nucleus of the ST (stb), there was a small decrease in the SP-positive fibers in the lateral hypothalamus (LH), a significant decrease in the SRIF-positive fibers in the lateroventral part of the anterior hypothalamic nucleus (lvAH), and a small decrease in the SRIF-positive fibers in the LH. These facts indicate that the origins of a number of SP- and SRIF-positive fibers are the Amc and that the amygdalofugal SP pathway in the ST innervates stb and LH and the amygdalofugal SRIF pathway in the ST projects to lvAH and LH.