Yuriko Kawai

Distribution of calcitonin gene-related peptide in the rat peripheral nervous system with reference to its coexistence with substance P

This immunocytochemical study, using a double-staining method, showed that calcitonin gene-related peptide-like immunoreactive structures are widely distributed in the peripheral nervous system and that many of them coexist with substance P-like immunoreactive structures in single sensory ganglion cells. Neurons positive for calcitonin gene-related peptide but negative for substance P were detected in sensory ganglia. These cells were large (about 30-45 micron in diameter); these primary sensory neurons containing calcitonin gene-related peptide can probably act independently of substance P. There were neurons containing calcitonin gene-related peptide without substance P in the pterygopalatine ganglion, although these cells were less numerous than in the sensory ganglia. In consecutive sections, calcitonin gene-related peptide-like structures occurred in thyroid parafollicular cells, which also contain calcitonin. This suggested that messenger RNA for producing calcitonin gene-related peptide is also present in the thyroid, and like calcitonin, calcitonin gene-related peptide may have a peripheral physiological role.

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.

Effect of calcitonin gene-related peptide on contraction of striated muscle in the mouse

We have found ultrastructurally calcitonin gene-related peptide-like immunoreactivity in the axon terminal within the synaptic trough of neuromuscular junction of the mouse. We determined, using pharmacological means, with a phrenic nerve-diaphragm preparation, that this peptide enhances muscle contraction during stimulation of the nerve fibers or direct stimulation of the muscle. This effect is probably brought about via the receptor for this peptide not the acetylcholine receptor.

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.

Ontogeny of somatostatin-containing neuron system of the rat cerebellum including its fiber connections: an experimental and immunohistochemical analysis.

Ontogeny of somatostatin (SRIF) neuron system of the rat cerebellum was investigated by means of the indirect immunofluorescent method. In addition, several operations were performed to provide the evidence of understanding the cerebellar SRIF innervation. SRIF-positive structures appeared in the primordium of the cerebellum of the fetus at gestational day 16 (14-15 mm embryos). From that time on, SRIF-positive structures identified in the cerebellum increased in number and the greatest number of SRIF-positive structures were detected in the cerebellum of the rat between newborn and 7-day-olds. Afterwards, these structures decreased remarkably in number and only a few if any SRIF-positive structures were observed in the cerebellum of the adult rats. These facts might suggest that SRIF might perform some important roles in the development of the cerebellum. The present study further elucidated that cerebellar SRIF was supplied by at least two types of sources; one was situated within the cerebellum and the other in the extracerebellar regions. The former one innervates mainly the vermis and partly the hemisphere, while the latter one projects mainly to the hemisphere and partly to the vermis.

Experimental and immunohistochemical studies on the cerebellar substance p of the rat: Localization, postnatal ontogeny and ways of entry to the cerebellum

With the indirect immunofluorescence technique, the localization (including the postnatal ontogeny) of substance P in the cerebellum, and the ways of entry of substance P-containing fibers into the cerebellum were explored. In the newborn rat cerebellum, dense fiber bands of axons with substance P-like immunoreactivity which can be traced to the lower brain stem are found. These fibers are also traceable to the developing granular cell layer. Two weeks after birth, however, substance P-containing structures seen in the cerebellum begin to decrease progressively and in the cerebellum of the adult rats, only a small amount of substance P-containing structures is observed. The present study established that substance P-containing fibers are mostly derived from extracerebellar substance P-containing cells and demonstrated the presence of three sites of entry of these substance P-containing fibers to the cerebellum, via (1) the inferior cerebellar peduncle, (2) the fasciculus uncinatus and (3) the middle cerebellar peduncle, respectively. Following deafferentation of the cerebellum, substance P-accumulating fibers are observed only ventral to the lesion (i.e. on the brain stem side), while in the cerebellum a remarkable decrease of substance P-containing fibers is seen and no substance P-accumulating fibers are found dorsal to the lesion (cerebellar side).