Biomedical subjects

H Kashiba

Publications and source records attributed to H Kashiba.

At least 19 recordsLinked to original sources

PubMed2001

Difference in binding by isolectin B4 to trkA and c-ret mRNA-expressing neurons in rat sensory ganglia.

The neurons labeled by isolectin B4 (IB4) in rat and mouse sensory ganglia are often regarded as non-nerve growth factor (NGF)-dependent and non-peptidergic neurons, but a considerable number of IB4-positive neurons in the dorsal root ganglion (DRG) are also shown to be immunoreactive to substance P (SP) and calcitonin gene-related peptide (CGRP), which are synthesized by NGF-dependent neurons. Therefore, we examined the relationships between the IB4-binding neurons and NGF/glial cell line-derived neurotrophic factor (GDNF)/GDNF-related proteins(GDNFs)-dependent neurons in rat DRGs by use of in situ hybridization histochemistry in serial sections. Of the DRG neurons, 42% and 22% were intensely and weakly labeled by IB4, respectively. The former neurons were small, and the latter varied in size. Of the trkA mRNA-expressing neurons, 29% and 57% were intensely and weakly labeled by IB4, respectively. On the other hand, 66% and 10% of the c-ret mRNA-expressing neurons were intensely and weakly labeled, respectively. The mRNA of somatostatin, another major neuropeptide in the sensory neurons, was exclusively expressed in the intensely IB4-labeled neurons. These findings suggest that many NGF-dependent and peptidergic sensory neurons are labeled by IB4 in rats.

Animals↗

PubMed2001

Histamine H1 receptor mRNA is expressed in capsaicin-insensitive sensory neurons with neuropeptide Y-immunoreactivity in guinea pigs.

Histamine H1 receptor mRNA-expressing sensory neurons in guinea pigs are unmyelinated and are not immunoreactive to substance P and calcitonin gene-related peptide (CGRP) [Mol. Brain Res. 66 (1999) 24], which are implicated in the nociceptive transmission of the primary sensory system. In this study, we examined whether these H1 receptor mRNA-expressing neurons are sensitive to capsaicin by using in situ hybridization histochemistry. Of lumbar dorsal root ganglion (DRG) neurons in control animals, 17% were positive for CGRP. In guinea pigs neonatally treated with capsaicin (50 mg/kg), few CGRP-immunoreactive neurons were seen in the DRGs. However, the percentages of H1 receptor mRNA-expressing neurons (15-20%) and the intensity of the mRNA signals in these neurons were not affected by neonatal capsaicin treatment. We also revealed the presence of both capsaicin-sensitive and insensitive neuropeptide Y (NPY)-immunoreactive neurons in the DRGs. These neurons were exclusively small. H1 receptor mRNA was expressed in NPY-immunoreactive neurons in naive guinea pig DRGs. These results suggest that H1 receptor mRNA is expressed in capsaicin-insensitive DRG neurons with NPY-immunoreactivity in guinea pigs.

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PubMed2001

[Primary sensory neurons expressing histamine H1-receptor mRNA].

Pharmacological studies have suggested that a subgroup of primary sensory neurons is responsive to histamine via the H1 receptor. However, which type of primary sensory neurons express H1 receptor is not known. We addressed this issue using in situ hybridization histochemistry with a cRNA probe for the guinea pig H1 receptor mRNA. H1 receptor mRNA was expressed in about 15-20% of the trigeminal and lumbar dorsal root ganglion (DRG) neurons, but none of the nodose ganglion neurons. The positive neurons in DRG were exclusively small in size and were labeled by isolectin B4, suggesting that these neurons have unmyelinated fibers. However, H1-receptor mRNA-expressing DRG neurons were not immunoreactive to substance P (SP) or calcitonin gene-related peptide (CGRP), which are implicated in the nociceptive transmission of the primary sensory system. Moreover, in guinea pigs neonatally treated with capsaicin (50 mg/kg), few CGRP-immunoreactive neurons were seen in DRG, but the percentage of H1-receptor mRNA-expressing neurons (15%-20%) and the intensity of the mRNA signals in these neurons were not affected by neonatal capsaicin treatment, suggesting that H1 receptor-expressing neurons are not sensitive to capsaicin. These findings suggest that H1-receptor-expressing neurons are involved in the transmission of a unique sensory modality such as itch. A marked increase in the number of mRNA-positive DRG neurons was observed 1-5 days after a crush injury of the sciatic nerve (3-4-fold of the control value). These neurons that turned mRNA-positive after the nerve crush were also mainly small-sized. The mRNA signals were detected in many peptidergic (SP/CGRP) neurons, in contrast to the normal condition. On the other hand, mRNA signals were decreased in the neurons that showed intense labeling in the normal condition. These results suggest that the gene expression of H1 receptors up-regulated in injured afferents may be involved in neuropathic pain.

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PubMed2000

Delayed expression of somatostatin mRNA in GDNFs-dependent rat sensory neurons during postnatal development.

Gene expression of somatostatin (SST) and preprotachykinin A (PPTA) in lumbar DRG neurons of postnatal developing rats was examined by in situ hybridization. SST mRNA signals were not seen in DRG neurons until postnatal day 1 to 7, and were detected in about 10% of DRG neurons of 2- and 8-week-old rats. The positive neurons expressed c-ret mRNA in 8-week-old rats. On the other hand, PPTA mRNA signals were constantly seen in about 30% of DRG neurons. This study demonstrates the differential expression patterns of SST and PPTA mRNAs in DRG neurons of developing rats.

Animals↗

PubMed1999

Up- and down-regulation of BDNF mRNA in distinct subgroups of rat sensory neurons after axotomy.

Anterograde transport of BDNF is enhanced by axotomy in the rat sciatic nerve. However, the changes in BDNF gene expression in dorsal root ganglion (DRG) neurons after axotomy are not known. We examined this issue using in situ hybridization histochemistry. BDNF mRNA was detected in 35-40% of DRG neurons (L5) of control rats. Most of these neurons are small. BDNF gene expression in these neurons was down-regulated after application of capsaicin to the sciatic nerve. Transection of the sciatic nerve induced the up-regulation of BDNF mRNA. The intensely labeled neurons were mainly large and immunoreactive for neuropeptide Y. These results suggest that up- and down-regulation of BDNF gene expression in distinct subgroups of rat DRG neurons are caused by damage to the peripheral nerve.

Afferent Pathways↗

PubMed1999

Gene expression of histamine H1 receptor in guinea pig primary sensory neurons: a relationship between H1 receptor mRNA-expressing neurons and peptidergic neurons.

Pharmacological studies have suggested that a subgroup of primary sensory neurons is responsive to histamine via the histamine H1 receptor. We addressed this issue using in situ hybridization histochemistry with a cRNA probe for the guinea pig H1 receptor gene. About 15% of the trigeminal and lumber dorsal root ganglion (DRG) neurons, but none of nodose ganglion neurons, were intensely labeled with this probe. The H1 receptor mRNA-positive neurons were exclusively small in size, and were demonstrated to give rise to unmyelinated fibers by ultrastructural analysis of isolectin B4-labeling. However, the H1 receptor mRNA-expressing DRG neurons were not immunoreactive to substance P (SP) and calcitonin gene-related peptide (CGRP). A marked increase in the number of mRNA-positive DRG neurons were observed 1-5 days after a crush injury of the sciatic nerve (3-4-fold of the control value). These neurons turned mRNA-positive after the nerve crush were also mainly small-sized. The mRNA signals were detected in many peptidergic (SP/CGRP) neurons, in contrast to the normal state. On the other hand, in the neurons which showed intense labeling in the normal condition, the mRNA signals were down-regulated. These results suggest that primary sensory neurons include two kinds of H1 receptor-expressing sensory neurons, one expressing H1 receptor mRNAs in the normal state and the other up-regulating the mRNAs following the peripheral nerve damage.

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PubMed1998

Glial cell line-derived neurotrophic factor and nerve growth factor receptor mRNAs are expressed in distinct subgroups of dorsal root ganglion neurons and are differentially regulated by peripheral axotomy in the rat.

We examined the colocalization of glial cell line-derived neurotrophic factor (GDNF) and nerve growth factor (NGF) receptor genes in rat dorsal root ganglion (DRG) neurons, and investigated the changes of the gene expression following sciatic nerve transection using in situ hybridization histochemistry. About 60% and 35% of the lumbar DRG neurons expressed c-ret and trkA, proto-oncogenes of the functional receptors for GDNF and NGF, respectively. Of the DRG neurons, however, only 9% was positive for both genes. A marked enhancement of the gene expression for GDNF receptor alpha (GDNFR alpha), which is a component of GDNF receptor, was observed in DRG neurons after sciatic nerve transection, but the percentage of c-ret mRNA-expressing neurons was not changed. The trkA mRNA-expressing neurons were decreased in number. These findings suggest that GDNF and NGF support distinct subgroups in intact DRG neurons, and that these receptor genes are differentially regulated when a peripheral nerve is injured.

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PubMed1997

Relationship between BDNF- and trk-expressing neurones in rat dorsal root ganglion: an analysis by in situ hybridization.

Brain-derived neurotrophic factor (BDNF) is synthesized in sensory neurones and suggested to operate on these same neurones by an autocrine mechanism, but it is unclear whether these neurones express the functional receptor (TrkB) for BDNF. We therefore examined the co-localization of BDNF and neurotrophin receptor mRNAs in adult rat dorsal root ganglion (DRG) neurones by in situ hybridization histochemistry. BDNF mRNA signals were detected in about 40-50% of DRG neurones (L4-5). Almost all the trkA mRNA-expressing neurones (95%) were positive for BDNF mRNA, while no trkB mRNA- and few trkC mRNA-expressing neurones displayed BDNF mRNA signals. These findings suggest that BDNF is mainly synthesized in the sensory neurones that are responsive to nerve growth factor but not to BDNF or neurotrophin-3. It is unlikely that BDNF is involved in an autocrine loop in sensory ganglia.

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PubMed1997

Systemic capsaicin in the adult rat differentially affects gene expression for neuropeptides and neurotrophin receptors in primary sensory neurons.

While systemic capsaicin in adult rats is known to reduce substance P and somatostatin in primary sensory nerves, it is still unknown if it also affects the production of these peptides at the genetic level. Therefore, we examined the effects of systemically administered capsaicin on the expression of the beta-preprotachykinin, gamma-preprotachykinin, somatostatin, calcitonin gene-related peptide, vasoactive intestinal polypeptide, galanin, neuropeptide Y and neurotrophin receptor family (trkA, trkB, trkC) genes in dorsal root ganglion neurons by in situ hybridization in adult rats. Nerve growth factor is thought to be involved in the regulation of some of these genes. In the control animals, beta-preprotachykinin, gamma-preprotachykinin, calcitonin gene-related peptide, somatostatin, trkA, trkB and trkC messenger RNAs were found in about 30%, 30%, 40%, 10%, 40%, 5% and 20% of the lumbar dorsal root ganglion neurons, respectively. The number of neurons expressing beta/gamma-preprotachykinin and calcitonin gene-related peptide messenger RNAs decreased to about 50% and 70% of the control values, respectively, six days after subcutaneous administration of capsaicin (950 mg/kg). Simultaneously, the number of trkA messenger RNA-expressing neurons also decreased to about 70% of the control level, while the number of neurons expressing trkB and trkC messenger RNAs was unaffected. On the other hand, vasoactive intestinal polypeptide and galanin messenger RNAs, but not neuropeptide Y messenger RNA, began to be expressed in about 10% of dorsal root ganglion neurons after administration of capsaicin, although their messenger RNAs were not detected in the controls. However, the expression of somatostatin messenger RNA was unaffected by the systemic administration of capsaicin. The somatostatin messenger RNA was not co-expressed with vasoactive intestinal polypeptide and galanin messenger RNAs in the sensory neurons of rats given capsaicin. Electron microscopic analysis revealed a few degenerating unmyelinated afferents in sural nerves of the treated rats. The number of small-sized dorsal root ganglion cells labeled with Fluoro-Gold, a retrograde-tracing dye which was injected into the sural nerve of the treated rats, decreased to half of the control number. Our results suggest that systemic administration of capsaicin in adult rats depresses the expression of beta/gamma-preprotachykinin, calcitonin gene-related peptide and trkA messenger RNAs, and induces expression of vasoactive intestinal polypeptide and galanin messenger RNAs in sensory neurons, which may be due to the capsaicin-induced degeneration of a subpopulation of sensory afferents. We also demonstrated that the regulation of somatostatin gene expression in mature sensory neurons is not affected by systemic capsaicin.

Animals↗

PubMed1996

Coexpression of preprotachykinin-A, alpha-calcitonin gene-related peptide, somatostatin, and neurotrophin receptor family messenger RNAs in rat dorsal root ganglion neurons.

Syntheses of substance P, somatostatin, and calcitonin gene-related peptide in sensory neurons have been suggested to be regulated by neurotrophic factors retrogradely transported from target tissues. In this study, we re-examined this idea by investigating the coexpression of neurotrophin receptor (trk family proto-oncogene) messenger RNAs, and preprotachykinin-A (a precursor peptide of substance P), alpha-calcitonin gene-related peptide and somatostatin messenger RNAs in lumbar dorsal root ganglion neurons by means of in situ hybridization histochemistry in rats. Approximately 35-40%, 5% and 15-20% of sensory neurons displayed signals for trkA, trkB, and trkC messenger RNAs, respectively. Approximately 28% of dorsal root ganglion neurons were positive for preprotachykinin-A messenger RNA, and were divided into two groups; those labeled strongly and those labeled weakly by in situ hybridization. All the strongly-labeled neurons (78% of preprotachykinin-A-positive cells) expressed trkA messenger RNA at the same time, while the weakly-labeled neurons did not. Thirty-seven per cent of dorsal root ganglion neurons expressed alpha-calcitonin gene-related peptide messenger RNA, and most of these neurons (84%) also expressed trkA messenger RNA. No or few preprotachykinin-A messenger RNA- and/or alpha-calcitonin gene-related peptide messenger RNA-expressing neurons were also positive for trkB or trkC messenger RNAs. Nine per cent of dorsal root ganglion neurons expressed somatostatin messenger RNA, and these neurons lacked all three trk messenger RNAs. Furthermore, most of these neurons (about 90%) showed positive, albeit weak, signals for preprotachykinin-A and alpha-calcitonin gene-related peptide messenger RNAs. The results suggest that expression of preprotachykinin-A and alpha-calcitonin gene-related peptide messenger RNAs is mediated by nerve growth factor via trkA receptor but not by brain-derived neurotrophic factor or neurotrophin-3, and that somatostatin gene transcription is not regulated by any member of the neurotrophin family in rat sensory neurons.

Animals↗

PubMed1996

Sensory afferent processing in multi-responsive DRG neurons.

The recent advance in molecular and neurobiological techniques disclosed the multi-responsive nature of DRG neurons. The survival, phenotype expression and electrical properties of these neurons are under the control of a variety of substances through their specific receptors. In pathological conditions, such as tissue inflammation or nerve injury, DRG neurons change their responsiveness through the dynamic reconstruction of their receptor system. This reconstruction is initiated by environmental stimuli. Thus the properties of polymodal nociceptors can be altered according to the environmental conditions. The whole story of this mechanism is not disclosed yet. In order to understand this mechanism, it is basically important to identify various receptor mRNAs in DRG neurons, precise localization of receptor proteins, site of synthesis and route of supply of ligands for these receptors.

Animals↗

PubMed1995

Coexpression of trk family members and low-affinity neurotrophin receptors in rat dorsal root ganglion neurons.

We examined the colocalization of trk family and low-affinity neurotrophin receptor (LANR) mRNAs by means of in situ hybridization histochemistry in rat dorsal root ganglion (DRG) neurons. About 35-40%, 5%, and 15-20% of DRG neurons were positive for trkA, trkB, and trkC mRNAs, respectively. Each member of the trk family was coexpressed with LANR. The trkB mRNA-expressing neurons essentially lacked trkA and trkC mRNAs, and most of trkA and trkC mRNAs-expressing neurons (85-90%) belonged to different subpopulations, suggesting that they are involved in different functions.

Animals↗

PubMed1994

Neuropeptide Y and galanin are coexpressed in rat large type A sensory neurons after peripheral transection.

Neuropeptide Y (NPY)-like immunoreactivity (IR) was observed in 20-30% of ipsilateral dorsal root ganglion (DRG) neurons (L4-5) after unilateral transection of rat sciatic nerve. Most of these neurons contained 200 kDa subunit of neurofilaments and galanin. Immunohistochemical analysis combined with retrograde tracing method demonstrated that NPY-IR was detected in cutaneous and muscular sensory, but not in visceral sensory neurons. These findings suggest that NPY coexists with galanin in injured large type A cells, which may innervate the mechanoreceptors in the skin and muscle, such as corpuscles of Meissner and Pacini, or muscle spindles.

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PubMed1993

Changes of vinculin and extracellular matrix components following blunt trauma to rat skeletal muscle.

The purpose of the present study was to determine changes in the localization of vinculin, Type IV collagen, laminin, and fibronectin during the damage-repair process in the skeletal muscle over several days following acute blunt trauma. A single blunt trauma to the gastrocnemius muscle of the rat was induced by applying 1.57 J of kinetic energy to one leg. By day 2 after trauma, vinculin had disappeared rapidly from the intracellular submembraneous region corresponding to the costamere, whereas Type IV collagen and laminin persisted around the degenerating myofibers (corresponding to the basement membrane). Fibronectin could be observed filling the widened endomysium and in the ruptured myofibers. In the early phase of muscle regeneration (day 5 after trauma), newly synthesized vinculin, Type IV collagen, and laminin formed ring-like structures around the regenerating myotubes. Fibronectin could also be observed filling the widened endomysium around the regenerating myotubes. Further, when small newly formed or splitted myofibers were observed in the areas of regenerating myofibers (day 10 after trauma), vinculin, Type IV collagen, laminin, and fibronectin were localized in specialized regions corresponding to the costamere, basement membrane, and endomysium. These results demonstrated that restoration of the costamere, basement membrane, and endomysium are essential to the maturation of regenerating myofibers after injury.

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PubMed1992

Co-localized but target-unrelated expression of vasoactive intestinal polypeptide and galanin in rat dorsal root ganglion neurons after peripheral nerve crush injury.

Expression of vasoactive intestinal polypeptide (VIP) and galanin in dorsal root ganglion (DRG) neurons is known to be induced by peripheral nerve injury. We investigated (1) whether VIP and galanin were co-expressed by DRG neurons and (2) whether such neurons innervated specified peripheral targets (visceral, cutaneous or muscular). An antibody to the 200 kDa neurofilament subunit (NF200) was used as a marker for large type-A cells in the DRG. VIP and galanin were respectively observed in 22% and 67% of DRG neurons at the L5 spinal level after crushing of the sciatic nerve. Most VIP-containing neurons were small type-B cells (about 90%) and approximately 95% of VIP-containing neurons also showed galanin-like immunoreactivity. Galanin was expressed by both large type-A and small type-B cells. Immunocytochemistry combined with a retrograde tracer revealed that about 70-80% of the small type-B cells in each sensory division displayed VIP-like immunoreactivity, and that most of the tracer-labeled neurons also expressed galanin. These findings suggest that the expression VIP and/or galanin in response to peripheral nerve crush injury is a property common to visceral, cutaneous and muscular sensory neurons.

Animals↗

PubMed1992

Axonal blockade induces the expression of vasoactive intestinal polypeptide and galanin in rat dorsal root ganglion neurons.

Nerve growth factor (NGF) undergoes retrograde transport from peripheral target organs, and has been recently reported to regulate the production of some neuropeptides in dorsal root ganglion (DRG) neurons. Therefore, to ascertain whether or not the expression of calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), and galanin was regulated by the retrograde transport of factors such as NGF, we carried out an immunocytochemical analysis using vinblastine as an axonal transport blocker and a monoclonal antibody to the NGF receptor (NGFR) as a marker of NGF-responsive neurons. The percentage of CGRP-containing DRG neurons (L5) was decreased by sciatic nerve transection or by the application of higher doses of vinblastine (0.3-0.6 mM) to the sciatic nerve. VIP and galanin were expressed in some DRG neurons after the application of a low dose of vinblastine (0.15 mM), which can block axonal flow without causing neuronal damage. The expression of these peptides was not affected by dorsal rhizotomy. About 70% of the CGRP-containing neurons also expressed NGFR, while most of the VIP-containing or galanin-containing neurons lacked NGFR. These findings indicate that the depletion of peripheral target-derived neurotrophic factor(s) other than NGF by axonal blockade may induce the gene expression of VIP and galanin.

Animals↗

PubMed1992

Expression of galanin in rat primary sensory afferents after moxibustion to the skin.

We examined the effects of moxibustion on primary sensory neurons in the skin of rats using immunocytochemistry combined with a fluorescent retrograde tracer dye, fluoro gold (FG). Galanin-like immunoreactive (IR) fibers were often observed in the dermis of treated skin at 18 hours after moxibustion, while such fibers were rarely detected in untreated (control) skin. Moreover, most of galanin-IR fibers also displayed substance P (SP)-like immunoreactivity. About 20-30% of the dorsal root ganglion (DRG) neurons labeled when FG was injected intradermally into the moxibustion-treated skin showed galanin-like immunoreactivity, while the proportion of FG-labeled neurons with such immunoreactivity was less than 10% in control DRGs. These results show that moxibustion induced galanin expression by primary sensory neurons containing SP. The possible functions of this peptide are discussed in relation to the effects of moxibustion.

Animals↗

PubMed1991

Acupuncture to the skin induces release of substance P and calcitonin gene-related peptide from peripheral terminals of primary sensory neurons in the rat.

We immunohistochemically examined the short term effects of electro-acupuncture (E-acupuncture) to the skin on substance P (SP)- and calcitonin gene-related peptide (CGRP) containing primary sensory neurons in the rat. Immunoreactivity to SP and CGRP in these neurons at the treatment site decreased after 30 min of E-acupuncture. These results suggest that E-acupuncture induces release of SP and CGRP from peripheral terminals of primary sensory neurons.

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