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Biomedical subjects

H Kiyama

Publications and source records attributed to H Kiyama.

At least 145 records · Page 8Linked to original sources
PubMed1993

Distribution of GAP-43 (B50/F1) mRNA in the adult rat brain by in situ hybridization using an alkaline phosphatase labeled probe.

GAP-43 (B-50,F1, pp46) is a calmodulin binding protein which is specific to the nervous system and also a substrate for the protein kinase C. Furthermore an enrichment of this protein in the growth cone and developmental brain indicate that this protein is related to nerve development, regeneration, and outgrowth. While its level dramatically decreases after the completion of synaptogenesis, the protein is still to some extent continuously expressed in certain regions of the mature brain. In order to clarify GAP-43 localization in mature normal rats, we investigated the distribution of GAP-43 mRNA in the rat central nervous system by using a non-radioisotopic in situ hybridization histochemistry. This method demonstrated GAP-43 mRNA expressing cells with high resolution. GAP-43 mRNA was more abundant in the forebrain than in the lower brainstem. Intense hybridization signal was observed in the mitral cells of olfactory bulb, cerebral cortex, CA3 region of hippocampus, diagonal band, substantia nigra, raphe nuclei, locus coeruleus, and dorsal motor nucleus of vagus. Weak to moderate hybridization signals were also widely expressed in thalamus, hypothalamus, and midbrain. Moreover, most noradrenergic, adrenergic, serotonergic, histaminergic, and caudal part of dopaminergic cells exhibited an intense GAP-43 mRNA signal. Thus, GAP-43 mRNA is abundantly expressed under normal conditions in the brain and may play an important physiological role particularly in the forebrain and in monoaminergic neurons supporting the findings that GAP-43 could be implicated in plasticity and monoamine release.

Alkaline Phosphatase↗
PubMed1993

Distribution of the substance P receptor (NK-1 receptor) in the central nervous system.

The overall distribution of mRNA for the substance P receptor (NK-1) was investigated in the rat brain by in situ hybridization histochemistry using synthetic oligonucleotide probes. The NK-1 positive cells were distributed throughout the brain. Among the positive cells an intense hybridization signal was observed in the basal ganglia and the dorsal tegmental areas. In the hippocampus, hypothalamus, midbrain, and medulla oblongata, weak to moderate positive signal were recorded in various areas. Comparing our results with those of previous workers on the localization of substance P using both immunohistochemistry and in situ hybridization various relationships between the ligand and its receptor localization were characterized. Generally the areas containing the abundant substance P immunoreactive (SP-IR) terminals were associated with NK-1 mRNA positive cells, some examples of this can be seen in the medial amygdaloid nucleus, the locus coeruleus, and the dorsal raphe nucleus. Contrary to these matched areas, we could not detect NK-1 mRNA positive cells in the substantia nigra, where there is a strong projection from the caudate putamen containing the substance P. Similarly the substantia gelatinosa in the trigeminal tract nucleus and the dorsal horn of the spinal cord where numerous SP-IR fibers are located, did not show abundant NK-1 mRNA positive cells. Only a few weakly labeled positive cells could be found.

Animals↗
PubMed1993

Region-specific expression of subunits of ionotropic glutamate receptors (AMPA-type, KA-type and NMDA receptors) in the rat spinal cord with special reference to nociception.

The present study attempted to explore the gene expression of the subunits (GluR1-4) of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type receptor, subunit (GluR5) of kainic acid (KA)-type receptor, NR1 [a subunit of N-methyl-D-aspartate (NMDA) receptors] and the possible glutamate-binding subunit of an NMDA receptor complex in the dorsal horn of the rat spinal cords using in situ hybridization histochemistry. These results were compared with those of the spinal motor neurons. Expression of the subunits of the AMPA-type receptor was also examined at the protein level using immunocytochemistry, with reference to the motor neurons. Although all the four subunits of the AMPA-type receptor were expressed throughout the dorsal horn, the pattern of expression was different according to the dorsal horn region and to the subunits. GluR2 showed the strongest expression in the dorsal horn. Huge numbers of strongly labelled cells formed a dense collection in lamina II and superficial parts of lamina III. Many neurons in lamina II and superficial parts of lamina III expressed GluR1 moderately. Scattered neurons moderately expressing GluR3 were also seen in these regions, while the expression of GluR4 was very low. Labelling of the dorsal horn neurons by the GluR5 probe was low, and NR1 probe and a glutamate-binding subunit of an NMDA receptor complex probe labelled them diffusely with low to moderate intensity. These findings show a close relationship between the glutamergic nociceptive primary afferent system and AMPA-type receptors in which GluR2 is especially highly expressed. The present study further showed that the expression pattern of the glutamate receptors in the spinal sensory neurons differs considerably from that of spinal motor neurons. Motor neurons very strongly express GluR3 and 4, while the expression of GluR2 and GluR1 is moderate and low, respectively. Expression of GluR5 is also low in the motor neurons. However, expression of NR1 and the glutamate-binding subunit of an NMDA receptor complex is very strong. These findings indicate that the subunit composition of the AMPA-type receptors regulating motor neurons is different from that of the AMPA-type receptors in the spinal sensory neurons, and that there are at least two kinds of glutamergic systems which regulate motor neurons: via AMPA-type receptors and via NMDA receptors.

Animals↗
PubMed1993

The differential expression patterns of messenger RNAs encoding non-N-methyl-D-aspartate glutamate receptor subunits (GluR1-4) in the rat brain.

The messenger RNA expression of non-N-methyl-D-aspartate glutamate receptor subunits (GluR1-4), considered alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid type, was investigated in rat brain by in situ hybridization histochemistry using oligonucleotide probes specific to each subunit sequence. GluR1-4 subunit messenger RNAs were expressed widely and abundantly throughout the CNS. However, the combination of expression pattern varied notably according to location. GluR2 messenger RNA was expressed most strongly and widely, with most areas except the Bergmann glia containing this messenger RNA. GluR4 messenger RNA was also present widely, although the expression level was low. However, we observed many areas which lacked or expressed very little GluR1 messenger RNA, such as some nuclei in the general motor system and auditory system. In addition, some nuclei in the hypothalamus and general somatosensory system lacked or expressed very little GluR3 messenger RNA. These results suggest that in the rat CNS non-N-methyl-D-aspartate receptors varied their composition according to the area where they were expressed, and that the combination pattern might be related to the functional role of neurons.

Animals↗
PubMed1993

Localization of oxytocin receptor messenger ribonucleic acid in the rat brain.

The expression of oxytocin receptor (OT-R) mRNA in the rat central nervous system was examined by in situ hybridization histochemistry using cRNA probe. Wide distribution of cells expressing OT-R mRNA was observed not only in the hypothalamus, but also in other regions. There were high levels of OT-R mRNA in the anterior olfactory nuclei, tenia tecta, olfactory tubercle, rostral most region of the frontal cortex, piriform cortex, layers 2 and 3 of the neocortex, bed nucleus of the stria terminalis, anterior medial preoptic nucleus (AV3V region), magnocellular preoptic nucleus, supraoptic nucleus, paraventricular hypothalamic nucleus, retrochiasmatic nucleus, ventromedial hypothalamic nucleus, paraventricular thalamic nucleus, central amygdaloid nucleus, medial amygdaloid nucleus, posterior cortical amygdaloid nucleus, amygdalohippocampal area, subiculum, prepositus hypoglossal nucleus, and dorsal motor nucleus of vagus. In most regions of the brain, our findings concurred with those obtained by receptor binding autoradiography using a ligand specific to OT. However, in the inferior olive nucleus, OT-R mRNA was not detected despite an abundance of binding sites showed by receptor binding autography. Despite this discrepancy OT appears to have central nervous system functions in addition to its hormonal functions.

Animals↗
PubMed1993

GAP-43 mRNA expression in facial motoneurons during regeneration: in situ hybridization histochemistry study using an alkaline phosphatase-labelled probe.

By means of in situ hybridization histochemistry using an alkaline phosphatase-labelled probe, we found an increase of mRNA for the growth-associated protein GAP-43 in rat facial motoneurons following axotomy of the facial nerve. After nerve resection, the increased level of GAP-43 mRNA was maintained for at least 8 weeks, while it returned to almost undetectable levels by 8 weeks after nerve crush injury. Thus, expression of GAP-43 mRNA in motor neurons paralleled the process of axonal regeneration. However, the increase of GAP-43 mRNA after resection was more pronounced than after crushing, in this way being different from the pattern of low-affinity nerve growth factor receptor mRNA expression.

Alkaline Phosphatase↗
PubMed1993

[Recent progress in the use of the techniques in molecular histochemistry].

Recent developments in molecular biology have made it possible to detect molecules such as mRNA and DNA in situ. In this report we focused on non-radioactive in situ hybridization histochemistry using the alkaline phosphatase-labeled oligodeoxynucleotide probe, which has recently been developed. We report the advantages of this method and some applications such as semiquantitative analysis and simultaneous demonstration of two mRNAs in a single section. In addition, vasoactive intestinal polypeptide (VIP) receptor mRNA was demonstrated in the lung as an application of in situ hybridization histochemistry in the field of thoracic diseases. We compared the localized of VIP mRNA-positive cells between normal animals and an animal mode of asthma.

Base Sequence↗
PubMed1992

Regional distribution of cells expressing glycine receptor alpha 2 subunit mRNA in the rat brain.

The alpha 2 subunit of the glycine receptor is expressed transiently in the rat brain during early development suggesting that this subunit may be replaced by the alpha 1 subunit in the adult brain. The expression of glycine receptor alpha 2 subunit mRNA was investigated in the 7-day-old rat brain by in situ hybridization histochemistry using oligonucleotide probes specific for this subunit. Neurons expressing alpha 2 subunit mRNA were found to be widely and abundantly distributed throughout brain. We compared the distribution of neurons expressing alpha 2 subunit mRNA with that of neurons expressing alpha 1 or beta subunit mRNA. In the lower brainstem, the location of the neurons expressing alpha 2 subunit mRNA was very similar to that of the neurons with alpha 1 or beta subunit mRNA. Neurons expressing beta subunit mRNA were widespread and numerous in the forebrain, where neurons with alpha 1 subunit mRNA were uncommon. The locations of the neurons labeled by the alpha 2 probe were very similar to those of the cells labeled by the beta probe. These findings suggest that the alpha 2 subunit is not only expressed by immature neurons containing the alpha 1 subunit, but is also common to most immature neurons having the glycine receptor. However, it should be noted that several neurons contained beta and/or alpha 1 subunit mRNA but lacked alpha 2 subunit mRNA, suggesting that the glycine receptor is heterogeneous in its composition during brain development.

Animals↗
PubMed1992

c-fos may code for a common transcription factor within the hypothalamic neural circuits involved in osmoregulation.

Conscious rats were given an intraperitoneal (i.p.) injection of isotonic or hypertonic saline, and killed 10-240 min later. In the hypothalamus of hypertonic saline-injected rats, c-fos-mRNA positive cells were mainly restricted to the supraoptic and paraventricular nuclei and to structures associated with the lamina terminalis of the third ventricle, including in particular the subfornical organ, the organum vasculosum of the lamina terminalis (OVLT) and the median preoptic nucleus. These structures comprise the proposed anterior hypothalamic 'osmoreceptor complex' for regulation of vasopressin release. The time course of the appearance and disappearance of c-fos mRNA signals was similar in all regions. Thus c-fos protein (Fos) may be a common transcription factor in the hypothalamic neural circuits involved in osmoregulation.

Animals↗
PubMed1992

Postnatal development of mRNA specific for a metabotropic glutamate receptor in the rat brain.

We examined the ontogenesis of a subtype of metabotropic glutamate receptors, termed mGluR1, which is linked to phosphoinositide metabolism, in various regions of rat brain during neonatal development. Northern blot analyses of mGluR1 mRNA indicated that mRNA increased monotonously or remained at plateau levels during the first 5 weeks after birth. In situ hybridization analyses supported this conclusion. The result is in contrast with the reported development of the activity in excitatory amino acid-stimulated phosphoinositide turnover during the same period. The latter increases during the first few weeks and then decreases sharply.

Animals↗
PubMed1992

The GABAA receptor gamma 1 subunit is expressed by distinct neuronal populations.

The distribution of GABAA receptor gamma 1 subunit was examined in the rat central nervous system using in situ hybridization histochemistry. The gamma 1 subunit was expressed in relatively limited areas compared to other subunits investigated previously. The brain regions strongly expressing this subunit were the septum, globus pallidus, bed nucleus of the stria terminalis, hypothalamic periventricular nucleus, supraoptic nucleus, medial and central nuclei of the amygdaloid complex, medial part of the medial geniculate body, substantia nigra pars reticulata, interpeduncular nucleus, lateral parabrachial nucleus, Purkinje cell layer of the cerebellum, and inferior olivary nucleus. This relatively limited expression implies a possible role of gamma 1 subunit in relation to some specific neuronal circuit.

Animals↗
PubMed1992

Localization of GABAA-receptor gamma 2-subunit mRNA-containing neurons in the rat central nervous system.

The localization of neurons containing mRNA of the gamma 2-subunit of GABAA receptors was examined in the rat central nervous system with in situ hybridization histochemistry using an oligonucleotide probe to the sequence of the gamma 2-subunit. Neurons containing the gamma 2-subunit mRNA were widely but unevenly distributed in the brain. The location of gamma 2-subunit mRNA-containing neurons differed from those containing alpha- or beta-subunits. According to our results brain regions can be divided into three categories: one containing only gamma 2-subunit, one containing mRNA of at least one subunit other than gamma 2 and one containing more than one other subunit but not the gamma 2-subunit. The distribution of strongly labeled nuclei partly coincided with that of glutamate decarboxylase, suggesting that the GABAA receptor gamma 2-subunit would be involved in an autoreceptive mechanism of the GABAergic transmission.

Animals↗
PubMed1992

GABAA receptor subunit messenger RNAs show differential expression during cortical development in the rat brain.

Developmental changes of the expression of various GABAA receptor subunits (alpha 1, alpha 3, alpha 4, beta 1-3, and gamma 2) were examined in the fetal rat cerebral cortex using in situ hybridization histochemistry. The subunits showed three main patterns of development. The alpha 1 subunit showed the first pattern, in which no expression was observed during embryonic development. The alpha 4 and beta 1 subunits showed the second pattern, in which expression was observed in both the undifferentiated neuroepithelium and the developing cortical layers. The alpha 3, beta 2, beta 3, and gamma 2 subunits showed the third pattern, in which expression was only seen in the developing cortical layers. These findings strongly suggest the following: (i) the alpha 1 subunit is involved in GABAergic transmission in the mature cerebral cortex; (ii) the alpha 4 and beta 1 subunits are involved in both the differentiation of the neuroepithelium and the development of the cortical plate, and (iii) the alpha 3, beta 2, beta 3, and gamma 2 subunits are involved in the development of the cortical plate. Subunits already expressed on embryonic day 13 (beta 1, beta 3, and gamma 2) appear especially likely to have a special role in neuronal development.

Animals↗
PubMed1992

Different postnatal development of cells expressing mRNA encoding neurotensin receptor.

In situ hybridization histochemistry revealed three different ontogenetic patterns of localized expression of the high-affinity type of neurotensin receptor mRNA in the developing rat brain: one comprises sites which showed transient expression of neurotensin receptor mRNA during the first postnatal week, the expression greatly decreasing thereafter (type I); another comprises sites at which there is a gradual increase in neurotensin receptor mRNA after birth, as there is in cell number and intensity, with advancing age, followed by a plateau (type II); the third comprises sites at which there is much expression of neurotensin receptor mRNA already at birth, and a slight decrease thereafter (type III). The cerebral cortex, except retrosplenial and entorhinal cortices, and the anterior dorsal thalamic nucleus exhibit the type I pattern, while the horizontal and vertical limbs of the diagonal band of Broca, magnocellular preoptic nucleus, substantia innominata, ventral part of the suprachiasmatic nucleus, medial habenular nucleus, ventral tegmental area and substantia nigra pars compacta exhibit the type II pattern. The tenia tecta, retrosplenial and entorhinal cortices exhibit the type III pattern. One of the most striking findings in this study was that the entire neocortex and most of the limbic cortex exhibit the type I pattern, i.e. neurotensin receptor mRNA is expressed transiently long before a neuronal network is established there. This suggests that neurotensin plays an important role in cortical development, other than its reported transmitter-like role in the adult.

Aging↗
PubMed1991

Postnatal ontogeny of cells expressing prepro-neurotensin/neuromedin N mRNA in the rat forebrain and midbrain: a hybridization histochemical study involving isotope-labeled and enzyme-labeled probes.

The postnatal ontogeny of cells expressing prepro-neurotensin/neuromedin N messenger RNA (prepro-NT/NN mRNA) in the rat forebrain and midbrain was investigated by in situ hybridization histochemistry. According to the pattern of expression during development, the cells which express prepro-NT/NN mRNA can be roughly divided into 2 groups. In type I cells, prepro-NT/NN mRNA expression reaches a maximum in terms of content during the postnatal period. After this early peak, cells of this type express the same or less prepro-NT/NN mRNA, reaching a plateau at an adult level that still contains a high level of expression. In type II cells, prepro-NT/NN mRNA appears during the postnatal period, and the expression decreases dramatically after the first postnatal week, being almost undetectable by a few weeks after birth. Type I cells were observed in the following areas: the piriform cortex, field CA1 of Ammon's horn, subiculum, vertical, and horizontal limbs of the diagonal band of Broca, intermediate part of the lateral septal nucleus, bed nucleus of the stria terminalis, medial preoptic area, lateral hypothalamus, caudal part of the caudate putamen, medial, cortical, and central amygdaloid nuclei, ventral tegmental area, deep mesencephalic nucleus, cuneiform nucleus, dorsal raphe nucleus, laterodorsal tegmental nucleus, parabrachial nucleus, and oral part of the pontine reticular nucleus. Cells of type II were observed in the following areas: the mitral cell layer of the olfactory bulb, rostral part of the caudate putamen, (anterior) cingulate cortex, and retrosplenial cortex (posterior cingulate cortex).

Aging↗
PubMed1991

Transient expression of neurotensin mRNA in the mitral cells of rat olfactory bulb during development.

The transient expression of neurotensin mRNA in the mitral cells of the rat olfactory bulb was demonstrated during the perinatal period using non-radioactive in situ hybridization in which an alkaline phosphatase labelled oligodeoxynucleotide probe was used. The relative cellular content of neurotensin mRNA signal was measured by use of a microdensitometer. Neurotensin mRNA positive cells were observed in the primordium of mitral cells on embryonic day 14 and their mRNA content increased gradually up to the day of birth. During the first postnatal week, the strength of their neurotensin mRNA signal decreased dramatically, and continued to decrease until in the adult olfactory bulb neurotensin mRNA was no longer detectable. This decrease of the neurotensin mRNA content coincided with a parallel decrease of neurotensin immunoreactivity observed in the lateral olfactory tract.

Animals↗
PubMed1991

The transient appearance of proneurotensin mRNA in the rat hypoglossal nucleus during development.

Using a sensitive non-radioactive in situ hybridization technique utilizing alkaline phosphatase labelled oligodeoxynucleotide neurotensin probe we have observed the transient expression of neurotensin mRNA in the rat hypoglossal nucleus. Neurotensin mRNA containing cells were first observed on embryonic day 14 in the primodium of the hypoglossal nucleus. The intensity of the mRNA signal increased during the perinatal period until a maximum signal was observed at birth. After this time, the intensity of the neurotensin mRNA signal decreased dramatically during the first postnatal week. Subsequently the mRNA signal decreased gradually during maturation until only a very weak signal was detected in the adult. The visualization of neurotensin mRNA containing cells in the hypoglossal nucleus has not previously been reported. The transient expression of NT mRNA containing cells prior to birth may suggest that neurotensin has a role in the development of this nucleus.

Aging↗