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ABSTRACT: It has been proposed that serotonin (5-HT) facilitates the chemosensory activity of the carotid body (CB). In the present study, we investigated mRNA expression and immunohistochemical localization of the 5-HT synthetic enzyme isoforms, tryptophan hydroxylase 1 (TPH1) and TPH2, and the 5-HT plasma membrane transport protein, 5-HT transporter (SERT), in the CB of the rat. RT-PCR analysis detected the expression of mRNA for TPH1 and SERT in extracts of the CB. Using immunohistochemistry, 5-HT immunoreactivity was observed in a few glomus cells. TPH1 and SERT immunoreactivities were observed in almost all glomus cells. SERT immunoreactivity was seen on nerve fibers with TPH1 immunoreactivity. SERT immunoreactivity was also observed in varicose nerve fibers immunoreactive for dopamine beta-hydroxylase, but not in nerve fibers immunoreactive for vesicular acetylcholine transporters or nerve terminals immunoreactive for P2X(3) purinoreceptors. These results suggest that 5-HT is synthesized and released from glomus cells and sympathetic nerve fibers in the CB of the rat, and that the chemosensory activity of the CB is regulated by 5-HT from glomus cells and sympathetic nerve fibers.
Histochemie 12/2012; · 2.59 Impact Factor
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ABSTRACT: Long-term hypoxia (days to weeks) increases phosphorylation of tyrosine hydroxylase (TH) at Ser31 and Ser40 in the carotid body (CB). In the present study, we examined the time course of TH phosphorylation at Ser31 and Ser40 in CB of rats exposed to short-term hypoxia (within 1 day) for 0-24h. Using immunoblotting, the signal intensities of both phosphorylated TH were more intense in CB of rats exposed to hypoxia for 6, 12, 18, and 24h than those of controls. Using immunohistochemistry, immunoreactive intensities of both phosphorylated TH were significantly more intense in glomus cells after rats were exposed to hypoxia for 6, 12, 18, and 24h than those of controls (p<0.05). These results show that phosphorylation of TH at Ser31 and Ser40 is increased in CB glomus cells by short-term hypoxia, suggesting that activation of TH via phosphorylation contributes to the facilitation of catecholamine biosynthesis in CB glomus cells at an early stage of hypoxia.
Respiratory Physiology & Neurobiology 11/2012; · 2.24 Impact Factor
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Journal of Histochemistry and Cytochemistry 09/2012; · 2.72 Impact Factor
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ABSTRACT: Under hypertension, it has been reported that the carotid body (CB) is enlarged and noradrenaline (NA) content in CB is increased. Therefore, it is hypothesized that morphological and neurochemical changes in CB are induced in hypertensive animal models. In the present study, we examined the morphological features and dopamine β-hydroxylase (DBH) immunoreactivity in CB of spontaneously hypertensive rats (SHR/Izm) and Wistar Kyoto rats (WKY/Izm). The CB of SHR/Izm was elongated in terms of the cross section of center and was enlarged in the reconstructed images compared with that of WKY/Izm, and the total volume of CB in SHR/Izm (0.048 ± 0.004 mm³) was significantly (p<0.05) increased compared with the value in WKY/Izm (0.032 ± 0.006 mm³). By immunohistochemistry, immunoreactivity for tyrosine hydroxylase in CB was mainly observed in glomus cells and the immunostaining properties were similar between WKY/Izm and SHR/Izm. On the other hand, DBH immunoreactivity was mainly observed in nerve fibers around blood vessels and observed in a few glomus cells in CB of WKY/Izm. The number of glomus cells with strong DBH immunoreactivity was increased in SHR/Izm compared with that in WKY/Izm. In conclusion, the present study exhibited the enlargement of CB as three-dimensional image and revealed the enhanced immunoreactivity for DBH of glomus cells in SHR/Izm. These results suggest that the morphology of CB is affected by the effect of sympathetic nerve and that the signal transduction from CB is regulated by NA in glomus cells under hypertensive conditions.
Autonomic neuroscience: basic & clinical 04/2012; 169(1):49-55. · 1.82 Impact Factor
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ABSTRACT: Staphylococcal enterotoxin A (SEA) is a leading causative toxin of staphylococcal food poisoning. However, it remains unclear how this toxin induces emesis in humans, primates, and certain experimental animals. To understand the mechanism of SEA-induced emesis, we investigated the behavior of SEA in the gastrointestinal (GI) tract in vivo using the house musk shrew (Suncus murinus). Immunofluorescence of GI sections showed that perorally administered SEA translocated from the lumen to the interior tissues of the GI tract and rapidly accumulated in certain submucosa cells. These SEA-binding cells in the submucosa were both tryptase- and FcεRIα-positive, suggesting these SEA-binding cells were mast cells. These SEA-binding mast cells were 5-hydroxytryptamine (5-HT)-positive, but the intensity of the 5-HT signal decreased over time compared to that of mast cells in the negative control. Furthermore, toluidine blue staining showed the number of metachromatic mast cells was decreased in the duodenal submucosa, suggesting that SEA binding induced degranulation and release of 5-HT from submucosal mast cells. These observations suggest that the target cells of SEA are submucosal mast cells in the GI tract and that 5-HT released from submucosal mast cells plays an important role in SEA-induced emesis.
FEMS Immunology & Medical Microbiology 12/2011; 64(3):392-402. · 2.44 Impact Factor
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ABSTRACT: The nasal cavity of adult Xenopus laevis (X. laevis) is composed of a series of three compartments: principal, middle, and inferior chambers. The principal chamber is lined with olfactory epithelium (OE), middle chamber with middle chamber epithelium (MCE), and inferior chamber with vomeronasal epithelium (VNE). In the present study, we examined developmental changes of lectin-binding patterns of the OE, MCE, and VNE by the use of four biotinylated lectins; DSL, DBA, PNA, and UEA-I. From Stage 59, just after the beginning of metamorphosis, the stainings of the free border for DBA and UEA-I were decreased in the OE and MCE, respectively, but the stainings of secretory granules (SGs) in the OE became intense. From Stage 63, sensory cells positive for DSL were increased in these three epithelia, and positive stainings for UEA-I and DBA increased in the SGs and Jacobson's glands (JGs), respectively. In addition, from 3 months after the end of metamorphosis, the stainings of sensory cells for PNA, DBA, and DSL changed in the OE, MCE, and VNE, respectively, and those of the SGs, Bowman's glands, and JGs also changed for several lectins. The present results showed that glycoconjugates expressed in three epithelia and their associated glands changed during and after the end of metamorphosis. These findings may indicate that the functional maturation of each epithelium depends not only on the maturation of sensory cells, but also on the maturation of the SGs in supporting cells of the OE and their associated glands after the end of metamorphosis.
The Anatomical Record Advances in Integrative Anatomy and Evolutionary Biology 03/2011; 294(5):839-46. · 1.47 Impact Factor
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ABSTRACT: Morphological development of sensory structures in the laryngeal mucosa of postnatal rats was observed by use of immunohistochemistry for protein gene-product 9.5 (PGP9.5). Moreover, expression changes of high affinity neurotrophin receptors, TrkA, TrkB and TrkC, and low affinity neurotrophin receptor p75(NTR) were examined to elucidate the relationship to morphogenesis. Intraepithelial nerve endings and parent axons of the laminar endings with immunoreactivity for PGP9.5 have already appeared in the rat on embryonic day 18 (E18) as well as solitary chemoreceptor cells in the glottic cleft. According to neurotrophin receptors, TrkA immunoreactivity were observed on and after postnatal week 3 (3W) in the nervous sensory structures, that is, free nerve endings, laminar endings and sub- and intragemmal plexuses of the taste buds. In the laminar endings, TrkC immunoreactivity was also observed on and after 3W. According to the laryngeal sensory cells, the solitary chemoreceptor cells were immunoreactive to TrkA, TrkB, and TrkC on and after postnatal day 3 (P3). In the taste buds in arytenoid region, taste cells were immunoreactive for TrkA, TrkB, and TrkC on and after 3W, P14, and 3W, respectively. Immunoreactivity for p75(NTR) was observed on the surface of taste cells on and after P9. The results of the present study suggest that sensory structures in the laryngeal mucosa were developed on perinatal days to involve respiratory reflex, and that neurotrophin receptors may take part in the regulation and maintenance of sensory structures.
The Anatomical Record Advances in Integrative Anatomy and Evolutionary Biology 03/2011; 294(4):694-705. · 1.47 Impact Factor
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ABSTRACT: Nitric oxide (NO) is a free radical and produced from L-arginine by nitric oxide synthase (NOS). Since NO is recently suggested to be involved in olfactory perception, the expression of eNOS, an isoform of NOS, was examined in the rat olfactory epithelium. The activity of NADPH-diaphorase was also examined as a marker of NOS. In the dorsomedial region of the nasal cavity, intensely positive reactions for NADPH-diaphorase were observed in the entire cytoplasm of sensory cells (olfactory cells). By immunohistochemistry, intensely positive reactions for eNOS were also found in the dorsomedial region of the nasal cavity. These reactions were observed on the free border of the olfactory epithelium. By immunoelectron microscopy, positive reactions for eNOS were found in the cilia of olfactory cells. In addition, in situ hybridization analysis of the olfactory epithelium revealed the expression of eNOS mRNA in the olfactory cells. These results indicate the presence of eNOS in the olfactory cells of the rat, and differential expression of eNOS in the olfactory epithelium depending on the regions of the nasal cavity. In addition, NO produced by eNOS may be involved in olfactory perception in the cilia of olfactory cells.
Journal of Veterinary Medical Science 11/2010; 73(4):423-30. · 0.85 Impact Factor
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ABSTRACT: The olfactory epithelium and the vomeronasal organ of the Japanese striped snake were examined by lectin histochemistry. Of the 21 lectins used in the study, all lectins except succinylated-wheat germ agglutinin (s-WGA) showed similar binding patterns in the vomeronasal receptor cells and the olfactory receptor cells with varying intensities. The binding patterns of s-WGA varied among individuals in the vomeronasal and olfactory receptor cells, respectively. Four lectins, Bandeiraea simplicifolia lectin-II (BSL-II), Dolichos biflorus agglutinin (DBA), Sophora japonica agglutinin (SJA), and Erythrina cristagalli lectin (ECL) stained secretory granules and the organelles in the olfactory supporting cells and did not stain them in the vomeronasal supporting cells. These results suggest that the glycoconjugate moieties are similar in the vomeronasal and olfactory receptor cells of the Japanese striped snake.
Journal of Morphology 10/2010; 271(10):1197-203. · 1.54 Impact Factor
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ABSTRACT: Neurochemical and morphological changes in the carotid body are induced by chronic hypoxia, leading to regulation of ventilation. In this study, we examined the time courses of changes in immunohistochemical intensity for tyrosine hydroxylase (TH) and cellular volume of glomus cells in rats exposed to hypoxia (10% O(2)) for up to 24 hr. Grayscale intensity for TH immunofluorescence was significantly increased in rats exposed to hypoxia for 12, 18, and 24 hr compared with control rats (p<0.05). The transectional area of glomus cells was not significantly different between experimental groups. The TH fluorescence intensity of the glomus cells exhibited a strong negative correlation with the transectional area in control rats (Spearman's rho = -0.70). This correlation coefficient decreased with exposure time, and it was lowest for the rats exposed to hypoxia for 18 hr (rho = -0.18). The histogram of TH fluorescence intensity showed a single peak in control rats. The peaks were gradually shifted to the right and became less pronounced in hypoxia-exposed rats, suggesting that a hypoxia-induced increase in TH immunoreactivity occurred uniformly in glomus cells. In conclusion, this study demonstrates that short-term hypoxia induces an increase in TH protein expression in rat carotid body glomus cells.
Journal of Histochemistry and Cytochemistry 09/2010; 58(9):839-46. · 2.72 Impact Factor
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ABSTRACT: In the carotid body (CB), it has been reported that the expressions of tyrosine hydroxylase (TH) mRNA and TH protein are enhanced by exposure to hypoxia. However, it is not known whether CO(2) affects the expression of TH in the CB. We examined the expression of TH mRNA and the immunoreactivity for TH in the CB of rats exposed to hypoxia (10% O(2)), hypercapnia (10% CO(2)) and hypercapnic hypoxia (10% O(2) and 10% CO(2)) for 2-24 h. The expression of TH mRNA in the CB was markedly enhanced in rats exposed to hypoxia for 4 h (6.6-fold), 6 h (6.0-fold) and 8 h (7.8-fold), and in rats exposed to hypercapnic hypoxia for 12 h (4.8-fold). The most intense TH immunoreactivity was observed in the CB from rats exposed to hypoxia for 12 and 24 h and to hypercapnic hypoxia for 24 h. The expressions of TH mRNA and the immunoreactivity for TH were not altered in the CB of rats exposed to hypercapnia. It is suggested that CO(2) does not affect TH expression in the CB, and that it inhibits hypoxia-enhanced TH expression.
Respiratory Physiology & Neurobiology 08/2010; 173(1):95-100. · 2.24 Impact Factor
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ABSTRACT: In the sensory ganglia, neurons are devoid of synaptic contacts, and ganglion neurons surrounded by one of glial cells, satellite cells. Recent studies suggest that neurons and satellite cells interact through neurotransmitters. In the present study, intracellular Ca(2+) ([Ca(2+)](i)) dynamics of neurons and satellite cells from one of viscerosensory ganglia, nodose ganglion (NG), were investigated by stimulation with glutamate and its agonist and/or the antagonist of the GABA(A) receptor bicuculline. In the specimens containing neurons with satellite cells, glutamate and a metabotropic glutamate receptor (mGluR) agonist t-ACPD evoked [Ca(2+)](i) increases in both neurons and surrounding satellite cells. Moreover, bicuculline also induced [Ca(2+)](i) increases in neurons and satellite cells. However, in the isolated neurons, bicuculline did not cause an increase in [Ca(2+)](i), suggesting that satellite cells are equipped with the ability to release GABA. In the neurons associated with satellite cells, the delay time until the onset of a response was shorter in the case of glutamate stimulation with bicuculline than that without bicuculline (107.3 +/- 93.4 vs. 231.8 +/- 97.0 s, p < 0.01). Furthermore, immunoreactivities for glutamate transporter, GLAST, and GABA transporter, GAT-3, were observed in both neurons and satellite cells of NG. In conclusion, the levels of [Ca(2+)](i) of NG neurons and surrounding satellite cells are increased by glutamate through at least mGluRs, and endogenous GABA modulates these responses; GABA inhibition is dependent on a close association between neurons and satellite cells. Such neuron-glia interaction in the nodose ganglion may regulate sensory information from visceral organs.
Histochemie 07/2010; 134(1):13-22. · 2.59 Impact Factor
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ABSTRACT: TREK belongs to a subfamily of tandem pore domain K+ channels, and consists of three subunits, TREK-1, TREK-2 and TRAAK. We examined the distribution of TREK-1, TREK-2 and TRAAK immunoreactive neurons in rat trigeminal sensory neurons. In the trigeminal ganglia, 31%, 43% and 60% of neurons were immunoreactive for TREK-1, TREK-2 and TRAAK, respectively. Mean sizes of TREK-1, TREK-2 and TRAAK immunoreactive trigeminal ganglion neurons were 447+/-185, 445+/-23 and 492+/-12 mm2, respectively. Furthermore, TREK channels were colocalized with cationic TRP channels, TRPV1, TRPV2 and TRPM8. TREK-1 immunoreactive neurons were colocalized with TRPV1 (57%), TRPV2 (11%) and TRPM8 (33%). TREK-2-immunoreactive neurons were colocalized with TRPV1 (33%), TRPV2 (9%) and TRPM8 (19%). TRAAK immunoreactive neurons were colocalized with TRPV1 (47%), TRPV2 (10%) and TRPM8 (22%). The present results revealed that TREK-1, TREK-2 and TRAAK channels colocalized with thermosensitive TRP channels in some small trigeminal ganglion neurons.
Neuroscience Letters 05/2009; 454(2):129-33. · 2.11 Impact Factor
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ABSTRACT: There are two functional pathways for the nasotrigeminal reflex: the spinal nucleus of trigeminal nerve (SPV) to the Kölliker-Fuse (KF) nucleus and the nucleus of solitary tract (NTS) to the lateral parabrachial nucleus (PBl). Although stimulation of the nasal mucosa by cool temperature induces respiratory depression, it is still unknown whether these nuclei are activated. In the present study, we examined the expression of Fos protein in rat brainstem neurons after nasal application of l-menthol, which is known to activate cold-sensitive nasal receptors. Application of l-menthol, but not paraffin oil, decreased the respiratory rate from 99.7+/-15.6 to 78.5+/-7.3 min(-1). Furthermore, a significantly higher density of Fos-immunoreactive cells was observed in the SPV and KF in the l-menthol rats than in the controls. In the SPV, the density of Fos-immunoreactive cells was highest at approximately 0.5mm rostral to the obex in both the l-menthol (48.5+/-11.5 cells/section) and paraffin oil (26.0+/-9.6 cells/section) groups. In the KF, the mean density of Fos-immunoreactive cells was highest at approximately 5.0mm rostral to the obex in both groups (l-menthol: 67.8+/-14.0 cells/section, control: 41.0+/-12.7 cells/section). The present study suggests that the SPV-KF pathway is important for the cold-induced respiratory depression.
Neuroscience Letters 05/2008; 435(3):246-50. · 2.11 Impact Factor
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ABSTRACT: The inwardly rectifying K+ channels, Kir1.1, Kir2.3 and Kir4.1-Kir5.1, are the candidate chemosensory molecules for CO2/H+. We determined the mRNA expression and immunohistochemical localization of these channels in the medulla oblongata of the rat. RT-PCR analysis revealed mRNAs of Kir1.1, Kir2.3, Kir4.1 and Kir5.1 were detected in the medulla. The immunoreactivities for Kir1.1, Kir2.3, Kir4.1, and Kir5.1 were observed in the medulla, and immunolabeling pattern was varied by the subunit. Immunoreactivities for Kir1.1 and Kir2.3 were observed in the nerve cell bodies and glial cells both in the chemosensory areas [nucleus tractus solitarius (NTS), nucleus raphe obscurus (RO), pre-Bötzinger complex (PreBötC)] and non-chemosensory area [hypoglossal nucleus (XII), inferior olive nucleus (IO)]. Kir4.1 immunoreactivity was observed in the glial cells and neuropil, especially in XII and IO. Kir5.1 immunoreactivity was observed in the nerve cell bodies in the XII, RO, and PreBötC, but not in the NTS or IO. In the NTS, a dense network of varicose nerve fibers showed immunoreactivity for Kir5.1. Our findings suggest that Kir channels may not act specific to the central chemoreception, but regulate the ionic properties of cellular membranes in various neurons and glial cells.
Journal of Veterinary Medical Science 04/2008; 70(3):265-71. · 0.85 Impact Factor
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ABSTRACT: Nociception in the trachea is important for respiratory modulation. We investigated the distribution, neurochemical characteristics, and origin of nerve endings with immunoreactivity for candidate sensor channels, TRPV1 and TRPV2, in rat trachea. In the epithelial layer, the intraepithelial nerve endings and dense subepithelial network of nerve fibers were immunoreactive for TRPV1. In contrast, TRPV2 immunoreactivity was observed mainly in nerve fibers of the tracheal submucosal layer and in several intrinsic ganglion cells in the peritracheal plexus. Double immunostaining revealed that some TRPV1-immunoreactive nerve fibers were also immunoreactive for substance P or calcitonin gene-related peptide, but neither neuropeptide colocalized with TRPV2. Injection of the retrograde tracer, fast blue, into the tracheal wall near the thoracic inlet demonstrated labeled neurons in the jugular, nodose, and dorsal root ganglia at segmental levels of C2-C8. In the jugular and nodose ganglia, 59.3% (70/118) and 10.7% (17/159), respectively, of fast blue-labeled neurons were immunoreactive for TRPV1, compared to 8.8% (8/91) and 2.6% (5/191) for TRPV2-immunoreactive. Our results indicate that TRPV1-immunoreactive nerve endings are important for tracheal nociception, and the different expression patterns of TRPV1 and TRPV2 with neuropeptides may reflect different subpopulations of sensory neurons.
Journal of Anatomy 01/2008; 211(6):775-83. · 2.37 Impact Factor
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ABSTRACT: We investigated the expression of three subunits of epithelial sodium channel (ENaC), alphaENaC, betaENaC and gammaENaC, in the nodose ganglion and laryngeal mucosa of rat by RT-PCR analysis and immunohistochemistry. PCR products of predicted size for alphaENaC, betaENaC and gammaENaC subunits were amplified from extract of nodose ganglion. Immunohistochemically, nodose ganglion neurons of medium to large diameter were immunoreactive for alphaENaC, betaENaC and gammaENaC. In the deep region of laryngeal submucosal layer, thick nerve fibers without varicosities were immunoreactive for alphaENaC, betaENaC and gammaENaC. In the laryngeal mucosa, terminal arborizations of the nerve endings, that immunoreacted for alphaENaC, betaENaC and gammaENaC were scattered in the lamina propria just beneath the epithelia of epiglottis and laryngeal vestibule. Double immunofluorescence with calretinin revealed that they were laminar nerve endings. Some thick nerve fibers near the laryngeal taste buds were also immunoreactive for betaENaC and gammaENaC, but negative for alphaENaC. In the larynx, ENaC channels may play important roles in mechanotransduction in the laminar endings and in the mechano- and chemotransductions in the taste bud-associated nerve fibers.
Neuroscience Letters 08/2006; 402(3):227-32. · 2.11 Impact Factor
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Advances in experimental medicine and biology 02/2006; 580:9-14; discussion 351-9. · 1.09 Impact Factor
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ABSTRACT: We succeeded in noninvasively analyzing the distribution of tetraploid (4n) cells in tetraploid<-->diploid (4n<-->2n) chimeric embryos by using enhanced green fluorescent protein (EGFP) transgenic (Tg) mouse embryos. We also evaluated whether this technique of analyzing 4n-cells in EGFP Tg 4n<-->2n chimeric embryos could be used to determine which characteristics of 4n-cells cause the death of 4n-embryos and restricted distribution of 4n-cells in 4n<-->2n-chimeric embryos after implantation. In our experiments, the distribution of 4n-cells in 4n<-->2n-embryos was normal until an embryonic age of 3.5 days (E3.5). With respect to morphological development, there were no differences between 4n-, diploid (2n), 4n<-->2n-, and diploid/diploid (2n<-->2n) chimeric embryos, but the number of cells in the tetraploid (4n) blastocyst was smaller than expected. This decrease in the number of cells may have caused cell death or reduced the rate of cell division in 4n-cells, and may have restricted the distribution of 4n-cells in 4n<-->2n-chimeric embryos. This study demonstrated the utility of EGFP transgenic mouse embryos for relatively easy and noninvasive study of the sequential distribution of cells in chimeric embryos.
Journal of Reproduction and Development 10/2005; 51(5):567-72. · 1.46 Impact Factor
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ABSTRACT: Immunoreactivity for vanilloid receptor subtype 1 (VR1) and its analogue vanilloid receptor-like protein 1 (VRL1) were examined in combination with immunoreactivity for substance P (SP) and calcitonin gene-related peptide (CGRP) in the rat larynx. VR1 and VRL1 immunoreactivity were observed in the intraepithelial free nerve endings, subepithelial nerve plexus and laryngeal epithelial cells. Most of VR1 immunoreactive nerves were also immunoreactive for SP or CGRP. VR1 immunoreactive intraepithelial nerve endings may be laryngeal nociceptors.
Autonomic Neuroscience 02/2005; 117(1):62-5. · 1.86 Impact Factor
