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E Senba

Publications and source records attributed to E Senba.

At least 91 records · Page 5Linked to original sources

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.

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Correlation between dendrodendritic synapses of adrenergic type and synaptically evoked hyperpolarization in the sympathetic ganglion of adult rats.

Intracellular recording and labeling with biocytin followed by electron microscopic observation were used to examine the nature and the morphological basis of a synaptically evoked hyperpolarization following spikes in the rat superior cervical ganglion neurons. A large hyperpolarization (the amplitude > 8 mV; the duration > 1 s following spikes) was elicited by repetitive stimulation of the preganglionic nerves in 8% of cells examined (n = 50). The alpha 2-adrenoceptor antagonist, yohimbine, reversibly attenuated the hyperpolarization, without affecting spikes. A nicotinic antagonist, hexamethonium, blocked both the hyperpolarization and spikes. Atropine had no effect of these responses. Electron microscopic observation of dendrites of these cells revealed that they received synaptic inputs of adrenergic type besides a cholinergic one from the preganglionic axons. Some dendrites served as presynaptic elements. These results strongly suggest that the hyperpolarization is an inhibitory postsynaptic potential and that this disynaptic response to the preganglionic stimulation is mediated mainly by two transmitters, acetylcholine and noradrenaline that are released from axodendritic and dendrodendritic synapses, respectively. We conclude that there appears to be an adrenergic inhibitory local circuit that modulates cholinergic transmission in the sympathetic ganglia.

Adrenergic alpha-1 Receptor Antagonists↗

Substance P induced by peripheral nerve injury in primary afferent sensory neurons and its effect on dorsal column nucleus neurons.

Using in situ hybridization and the retrograde tracer, Fluorogold, we examined the expression of preprotachykinin (PPT) mRNA in the rat dorsal root ganglion neurons projecting to the gracile nucleus. Seven days after unilateral sciatic nerve transection, some medium- to large-sized neurons in the rat dorsal root ganglia projecting to the gracile nucleus express PPT mRNA, whereas very few gracile nucleus-projecting neurons on the contralateral side express PPT mRNA. Immunohistochemistry revealed an increase in substance P (SP) immunoreactivity in the gracile nucleus and large myelinated fibers in the dorsal root 2 weeks after unilateral sciatic nerve transection. The results suggest that medium to large DRG cells that project to the gracile nucleus express PPT mRNA de novo in response to peripheral nerve injury, and increased SP is transported to the gracile nucleus through large myelinated fibers. To determine whether the increased SP might affect the excitability of the gracile nucleus neurons postsynaptically, Fos expression after electrical stimulation of the injured sciatic nerve was examined. Multiple injections of the NK-1 receptor antagonist, CP-96,345, suppressed stimulus-induced Fos expression in gracile nucleus neurons including thalamic relay neurons. The inactive enantiomer, CP-96,344, had no effect on stimulus-induced Fos expression. These data indicate that the de novo synthesized SP in the lesioned primary afferent neurons may be involved in an augmentation of excitability in the dorsal column-medial lemniscus sensory pathway. This hyperexcitability may play a role in the pathogenesis of abnormal neuropathic sensations following peripheral nerve injury.

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Differential regulation of IEGs in the rat PVH in single and repeated stress models.

Various kinds of stressors induce immediate early genes (IEGs) in discrete brain regions. We recently reported the reduced response of Fos expression in the hypothalamic paraventricular nucleus (PVH) when rats are repeatedly exposed to immobilization (IMO) stress. In this study, using in situ hybridization histochemistry, we further extended the research to other IEGs, and the results showed that prior exposure to IMO for 6 days suppressed the induction of fosB, junB and NGFI-B, but NGFI-A, mRNAs in response to a challenge IMO on day 7, suggesting that repeated stress has different effects on the transcription of NGFI-A and the other IEGs, in the PVH.

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Axotomy induces preprotachykinin gene expression in a subpopulation of dorsal root ganglion neurons.

The distribution of dorsal root ganglion (DRG) cell sizes that show changes in preprotachykinin (PPT) gene expression and substance P (SP) levels following axotomy was examined using RNA blot analysis, in situ hybridization histochemistry, and immunocytochemistry. PPT mRNA was induced in medium-sized (1,000-2,000 microns 2) and large-sized (> 2,000 microns 2) cells in the DRG after axotomy. There was a 165% increase in the number of labeled cells after sciatic transection and a 260% increase after spinal nerve transection which results in axotomy of all the cells in the ganglion. The further increase after spinal nerve transection suggests that the induction occurred in axotomized neurons. PPT mRNA label was also present in a reduced number of small (< 1,000 microns 2) cells after axotomy. SP immunoreactivity was also induced in medium- and large-sized cells and reduced in small-sized cells. Our findings suggest that the expression of the PPT gene and SP is differentially regulated in different subpopulations of DRG neurons after axotomy and is consistent with the hypothesis that tachykinins may be important in both sensory transmission and regeneration.

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Repeated stress reduces the subsequent stress-induced expression of Fos in rat brain.

Repeated stress is known to potentiate the CNS response to subsequent stress. Various stressful stimuli can induce Fos expression in discrete regions of the brain, such as the lateral septum, the hypothalamic paraventricular nucleus and the locus coeruleus. We investigated by immunohistochemistry the effect of the stress of repeated immobilization on Fos expression in those regions of the brain in adult male rats. Six daily immobilizations suppressed the expression of Fos in all regions when immobilization was subsequently applied, suggesting that Fos does not play a major role in potentiating the stress response under repeated stressed conditions.

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Differential expression of fos family and jun family mRNAs in the rat hypothalamo-pituitary-adrenal axis after immobilization stress.

We aimed to clarify the regulatory mechanism of the hypothalamo-pituitary-adrenal axis that plays key roles in initiating stress responses, as well as the roles of immediate early genes in this process. We investigated the stress-induced activation of fos and jun family proto-oncogenes by means of in situ hybridization histochemistry. Immobilization stress induced c-fos and jun B mRNAs in the parvocellular region of the hypothalamic paraventricular nucleus, the anterior and intermediate lobes of pituitary, and in the adrenal gland after 7 min of immobilization, although no c-fos or jun B mRNAs were detected in these and other organs in control rats. The levels of these mRNAs peaked after 30-60 min of immobilization, then declined. A low level of fos B mRNA appeared at 15-30 min and peaked after 60-90 min. On the contrary, c-jun and jun D mRNAs were constitutively expressed in the paraventricular nucleus and adrenal cortex. These findings indicate that the members of the fos and jun family proto-oncogenes play different roles in the transcriptional regulation of genes involved in the hypothalamo-pituitary-adrenal axis, and that monitoring immediate early genes is a useful method for following stress-induced cellular responses in the neuro-endocrine system.

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The expression of neuropeptides and their mRNAs in the trigeminal mesencephalic nucleus following masseteric nerve transection.

By in situ hybridization and immunohistochemistry, we examined the expression of neuropeptides such as neuropeptide Y (NPY), galanin (Gal), substance P (SP), vasoactive intestinal polypeptide (VIP) and their mRNAs in the rat mesencephalic trigeminal nucleus (Mes5) following masseteric nerve transection. On the side contralateral to the nerve transection, none of the peptides examined were labeled in Mes5 cell bodies. However, on the side ipsilateral to the lesion, NPY, Gal and preprotachykinin (PPT) mRNAs appeared in Mes5 cell bodies. Double labeling for mRNAs by in situ hybridization and retrograde tracer fluoro-gold (FG) revealed that almost all (96-97%) the FG-labeled neurons which were cut expressed NPY and Gal mRNAs, whereas less neurons (87%) expressed PPT mRNA. NPY and Gal-like immunoreactivities were detected in Mes5 cell bodies ipsilateral to the axotomy. The results suggested that these neuropeptides play roles in adaptive processes after peripheral nerve injury in Mes5 neurons as they are thought to do so in dorsal root ganglion neurons.

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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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Projections of tachykinin- and glutaminase-containing rat retinal ganglion cells.

Glutamate (Glu) and the tachykinin substance P (SP) have been proposed as neurotransmitters or neuromodulators of the retinal projection to the brain. In the present study, we demonstrate that tachykinin-like (TK) immunoreactivity (IR) accumulates in rat retinal axons following electrical lesions to the optic tract, indicating that SP is conveyed in the optic nerve to its central targets. In addition, we show that eye enucleation causes a dramatic decrease in TK-IR fibers in the pretectal olivary nucleus (PON), but not in other retinorecipient nuclei of the thalamus and the midbrain, and that Fluorogold injected into the pretectum is retrogradely transported to the somata of TK-IR retinal ganglion cells (RGCs), indicating an important projection of TK-IR RGCs to the PON. We also show that most rat RGCs are labeled with antibodies against phosphate-activated glutaminase, an enzyme considered to generate the transmitter pool of glutamate. Unlike TK-IR fibers, phosphate-activated glutaminase-IR structures disappear in most retinorecipient nuclei following eye enucleation. The present results give neuroanatomical support to the idea that glutamate is a neurotransmitter in the retinal projection and suggest an important role for TK-IR RGCs in the relay of visual information to the PON.

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Quantification of axotomy-induced alteration of neuropeptide mRNAs in dorsal root ganglion neurons with special reference to neuropeptide Y mRNA and the effects of neonatal capsaicin treatment.

Alteration in mRNA expression in dorsal root ganglia (DRG) neurons encoding 5 neuropeptides was quantitatively compared in normal rats and in those neonatally treated with capsaicin, a selective neurotoxin which destroys a subpopulation of DRG neurons with unmyelinated axons. Adult rats received a unilateral transection of the sciatic nerve and were killed 7 days later. Oligonucleotide probes specific for the genes encoding neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), galanin (GAL), somatostatin (SOM), and calcitonin gene-related peptide (CGRP) were used for in situ hybridization and RNA blot analysis. Following the nerve cut, RNA blot analysis demonstrated a dramatic induction of NPY, VIP, and GAL mRNA levels from the undetectable constitutive level of expression. Conversely, CGRP and SOM mRNAs, which are constitutively expressed, were reduced 55% and 70%, respectively, following the nerve cut. A unimodal size distribution for neurons expressing NPY mRNA was determined, with a mean cross-sectional area of 1700 microns2 representing 24.4% of DRG neurons ipsilateral to the nerve cut. Neurons expressing VIP mRNA were mainly small sized, with a cross-sectional area of approximately 700 microns2, while those expressing GAL mRNA were both small (approximately 700 microns2) and medium (approximately 1,300 microns2) sized. The percentages of neurons expressing VIP or GAL mRNA were 19.9% and 33.7%, respectively. In neonatal capsaicin-treated rats, there was a 10% reduction in neurons expressing NPY mRNA, a 37% reduction for VIP, and a 27% for GAL mRNA compared to vehicle-treated rats after nerve cut. Capsaicin-sensitive neurons comprised 37% of CGRP neurons and 83% of SOM neurons. These observations suggest that NPY is primarily induced in myelinated primary afferent neurons, while VIP and GAL mRNA induction occurs in a mixed population, a sizeable percentage of which has unmyelinated axons. Additionally, SOM mRNA expression is associated mainly with unmyelinated primary afferents.

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Principal neurons as local circuit neurons in the rat superior cervical ganglion: the synaptology of the neuronal processes revealed by intracellular injection of biocytin.

To analyze the local circuitry of the sympathetic ganglion, the synaptic relations of the neuronal processes of the principal neurons in the rat superior cervical ganglion were investigated by correlated light and electron microscopy combined with intracellular injection of biocytin. Intracellular iontophoresis of biocytin followed by avidin-biotinylated horseradish peroxidase cytochemistry allowed complete visualization of the neuronal processes of the principal neurons. The stained principal neurons have a single process (axon), which leaves the ganglion, and several intraganglionic processes (dendrites), some of which show specific terminal arborizations. Some terminals of the dendritic collaterals formed pericellular plexuses or intercellular glomerular plexuses. Electron microscopically, the dendrites and their collaterals contain numerous small vesicles. Synaptic membrane specializations were observed between the stained dendritic collaterals and unlabeled neurites. These may be both preganglionic axon terminals and processes of principal neurons. The likely direction of neurotransmission often could not be determined because of the bidirectional synaptic structures. Our findings show that the dendritic collaterals of principal neurons appear to make both post- and presynaptic contacts with both the principal neurons and the preganglionic axons. It is suggested that the principal neurons might participate in local circuits involving not only preganglionic axons but also neighboring principal neurons.

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Stress-induced c-fos expression in the rat brain: activation mechanism of sympathetic pathway.

To clarify which brain regions are activated by stress, we used expression of the proto-oncogene, c-fos, as a marker. An increased number of neurons expressing Fos-like immunoreactivity in their nuclei was observed in discrete brain regions, such as the lateral septum, midline nuclei of the thalamus, paraventricular hypothalamic nucleus, brain stem catecholaminergic, and serotonergic neurons, in response to pain or immobilization stress. Distribution patterns of Fos-like immunoreactive neurons were quite similar in animals subjected to pain or immobilization. Whether or not neurons projecting to the spinal cord to activate the sympathetic pathway express Fos-like immunoreactivity was examined by means of fluorescent double-labeling using fluoro-gold (FG) as a tracer. In the PVH, Fos-immunoreactive neurons were localized in the dorsal medial parvocellular part, although those projecting to the spinal cord were localized dorsally and ventrally. Less than 1% of Fos-positive neurons in this nucleus was colocalized with FG. Among fos-like immunoreactive catecholaminergic and serotonergic neurons in the brain stem, those in the A5 region and raphe pallidus projected to the spinal cord.

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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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Histamine-caused itch induces Fos-like immunoreactivity in dorsal horn neurons: effect of morphine pretreatment.

It is well known that cutaneous pain causes c-fos gene expression in the dorsal horn neurons. The present study examined whether or not an itching sensation had the same effect on these neurons. In order to produce an itching sensation, cotton balls soaked in histamine solutions of different concentrations were applied to the lower limbs of rats and then the treated area was scratched with the tip of a pair of forceps. After 2 h, the number of neurons showing Fos-like immunoreactivity was significantly increased in a dose-dependent manner in the ipsilateral dorsal horn when compared to saline-treated and scratched controls. When scratching with the forceps was omitted, the number of these neurons was reduced to one-third, but was still significantly higher than in controls treated only with saline. Expression of Fos-like immunoreactivity in these animals was markedly reduced by morphine pretreatment (10 mg/kg, i.p.), suggesting that the transmission of both itch and pain is blocked by morphine.

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