Peptide 19 in the rat vagal and glossopharyngeal sensory ganglia

Peptide 19 (PEP 19) is a 7.6-kDa polypeptide which binds to calmodulin and inhibits calcium-calmodulin signaling. In this study, PEP 19-immunoreactivity (PEP 19-IR) was examined in the rat vagal and glossopharyngeal sensory ganglia. Twenty-nine percent, 59%, and 41% of sensory neurons contained PEP 19-IR in the jugular, petrosal, and nodose ganglia, respectively. These neurons were of various sizes (jugular, mean +/- SD = 635.8 +/- 392.6 microm2, range = 105.9-1695.9 microm2; petrosal, mean +/- SD = 370.9 +/- 228.5 microm2, range = 57.7-1662.7 microm2; nodose, mean +/- SD = 380.5 +/- 157 microm2, range = 87.5-950.4 microm2) and scattered throughout these ganglia. Double immunofluorescence method revealed that PEP 19-IR neurons which had parvalbumin-IR were rare in the ganglia (jugular, 4%; petrosal, 10%; nodose, 8%). PEP 19-IR neurons which contained calbindin D-28k were abundant in the petrosal (20%) and nodose (22%) ganglia but not in the jugular ganglion (8%). Retrograde tracing method indicated that many PEP 19-IR neurons projected to the circumvallate papilla and soft palate. In the soft palate, taste buds were innervated by PEP 19-IR nerve fibers. The present study suggests that PEP 19-IR neurons include chemoreceptors in the vagal and glossopharyngeal sensory ganglia.     

Delta-opioid receptor-immunoreactive neurons in the rat cranial sensory ganglia

Immunohistochemistry for delta-opioid receptor (DOR) was performed on the rat cranial sensory ganglia. The immunoreactivity was detected in 16%, 19% and 11% of neurons in the trigeminal, jugular and petrosal ganglia, respectively. The nodose ganglion was devoid of such neurons. DOR-immunoreactive (IR) neurons were mostly small to medium-sized (trigeminal, range = 62-851 microm(2), mean +/- SD = 359 +/- 175 microm(2); jugular, range = 120-854 microm(2), mean +/- SD = 409 +/- 196 microm(2); petrosal, range = 167-1146 microm(2), mean +/- SD = 423 +/- 233 microm(2)). Double immunofluorescence method revealed that all DOR-IR neurons were also immunoreactive for calcitonin gene-related peptide. The cutaneous and mucosal epithelia in the oro-facial region, tooth pulp, taste bud and carotid body were innervated by DOR-IR nerve fibers. In the brainstem, IR nerve terminals were located in the superficial medullary dorsal horn and dorsomedial part of the subnucleus oralis as well as the solitary tract nucleus. The present study suggests that DOR-IR neurons may be associated with nociceptive and/or chemoreceptive function in the cranial sensory ganglia.     

Neurocalcin-immunoreactive neurons in the petrosal ganglion innervate the taste bud

The distribution and origin of neurocalcin-immunoreactive (NC-ir) nerve fibers in the taste bud and carotid body were examined by an immunofluorescence method. In the circumvallate papilla of the tongue, NC-ir nerve fibers made subepithelial nerve plexuses and occasionally penetrated the taste bud. However, the carotid body was devoid of ir nerve fibers. In the petrosal ganglion, 32% of neurons were immunoreactive for NC. Such neurons were mostly medium-sized to large, and scattered throughout the ganglion. In the superior cervical and intralingual ganglia, numerous ir varicose fibers surrounded postsynaptic neurons. However, NC-ir could not be detected in cell bodies of these neurons. The retrograde tracing method indicated that NC-ir petrosal neurons innervated taste buds in the circumvallate papilla. NC-ir neurons may have a gustatory function in the petrosal ganglion.     

Peptide-containing Neurons Projecting to the Tongue of the Rat: Retrograde Tracing and Immunocytochemistry

The origin and neuropeptide content of nerve fibres in the rat circumvallate papilla was studied by retrograde tracing in combination with immunocytochemistry. An injection of the retrograde tracer True Blue into the circumvallate papilla resulted in the appearance of labelled nerve cell bodies in the superior cervical, the stellate, the thyroid, the nodose, the jugular, the petrosal, the otic, the trigeminal and the dorsal root ganglia at level C2. Most of the True Blue-labelled nerve cells in the superior cervical ganglia contained neuropeptide Y. The majority of labelled cell bodies in the thyroid ganglia contained vasoactive intestinal peptide. In the jugular and trigeminal ganglia, the majority of the labelled nerve cell bodies stored calcitonin gene-related peptide. A small number of neurons in the medial reticular formation of the central nervous system was labelled. Tracer injections deep into the tongue tissue beneath the circumvallate papilla gave rise to True Blue-labelled neurons in the hypoglossal nucleus.     

Co-expression of VRL-1 and calbindin D-28k in the rat sensory ganglia

The co-expression of vanilloid receptor 1-like receptor (VRL-1), a newly cloned capsaicin-receptor homologue, with calbindin D-28k was examined in the rat sensory ganglia. The co-expression was rare in the dorsal root, trigeminal and jugular ganglia and abundant in the petrosal and nodose ganglia. In the dorsal root ganglion, none of VRL-1-immunoreactive (ir) neuron co-expressed calbindin D-28k-immunoreactivity (ir). Of the VRL-1-ir neurons, 9 and 5% showed calbindin D-28k ir in the trigeminal and jugular ganglia, respectively. On the other hand, 35 and 63% of VRL-1-ir neurons in the petrosal and nodose ganglia, respectively, co-expressed these substances. The retrograde tracing method indicated that petrosal neurons which co-expressed VRL-1-and calbindin D-28k-ir innervated taste buds in the circumvallate papilla. The present findings may suggest that VRL-1 is associated with chemosensory functions in visceral sensory neurons.     

Ca2+/calmodulin-dependent protein kinase II in the rat cranial sensory ganglia

Immunohistochemistry for Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) was performed on the rat cranial sensory ganglia. More than one half of neurons was immunoreactive for the enzyme in the trigeminal (60%), jugular (70%), petrosal (55%) and nodose ganglia (63%). These neurons were mainly small to medium-sized. The co-expression study demonstrated that one half of CaMKII-immunoreactive (ir) neurons was also immunoreactive for calcitonin gene-related peptide (CGRP) or the vanilloid receptor subtype 1 (VR1) in the trigeminal, jugular and petrosal ganglia. In the nodose ganglion, CaMKII-ir neurons were mostly devoid of CGRP-immunoreactivity (ir) (8.2%) whereas the co-expression with VR1-ir was common among such neurons (72%). In the facial skin, nasal mucosa and palate, the epithelium and taste bud were innervated by CaMKII-ir nerve fibers. In addition, the retrograde tracing study demonstrated that 39.6% and 44.8% of trigeminal neurons which were retrogradely traced with fluorogold from the facial skin and nasal mucosa exhibited CaMKII-ir. Forty-six percent of petrosal neurons which innervated the soft palate were immunoreactive for the enzyme.     

Calretinin-immunoreactivity in vagal and glossopharyngeal sensory neurons of the rat: distribution and coexistence with putative transmitter agents.

Immunoreactivity for the calcium binding protein, calretinin (calretinin-ir), was demonstrated in cell bodies of vagal and glossopharyngeal sensory ganglia (jugular, petrosal, and nodose ganglia) and in associated nerve fibers. In the jugular and petrosal ganglia, many calretinin-ir neurons were also immunoreactive for calcitonin gene-related peptide and substance P. In the nodose ganglion, most of the calretinin-ir neurons lacked these peptides. None of the calretinin-ir neurons in these ganglia were also immunoreactive for tyrosine hydroxylase.     

Brn-3a deficiency increases tyrosine hydroxylase-immunoreactive neurons in the dorsal root ganglion

Immunohistochemistry for tyrosine hydroxylase (TH) was performed on the dorsal root ganglia (DRG) in wild-type, heterozygous and Brn-3a knockout mice at embryonic day 18.5. TH-immunoreactive (-IR) neurons were detected in the DRG of wild-type and heterozygous mice, but their proportion was greatly increased by the loss of Brn-3a function (wild-type and heterozygot, 8.4%; knockout, 20.9%). IR neurons were of various sizes in wild-type (mean+/-S.D.=118.1+/-55.4 microm2, range=26.6-306.3 microm2) and heterozygous mice. In the knockout mice, however, TH-IR neurons were mostly small (mean+/-S.D.=68.2+/-34.3 microm2, range=11.8-166.8 microm2). The present study suggests that Brn-3a may normally suppress TH expression in many small DRG neurons but activate TH expression in large DRG neurons.     

An in vitro model for the study of the role of innervation in circumvallate papillae morphogenesis.

The following study was done to demonstrate the reliability of an in vitro model for use in the study of early events and the role of innervation in mouse circumvallate papillae development. Gestational day (gd)-11 fetuses were partially dissected to produce explants that included the mandibular, hyoid, third and fourth branchial arches and their ganglia. In ganglionectomized explants, the nodose ganglia and either the geniculate, petrosal or both ganglia were removed. Explants were cultivated in roller tube culture for 24, 48, 72, and 96 h of culture and examined for the presence of papillary structures. Innervation was verified by immunostaining for neural cell adhesion molecule (NCAM). In all control explants, circumvallate papillae had formed by 72 h in culture. These papillae were innervated by fibers originating in petrosal or nodose ganglia, although, in a small number, fibers from the geniculate also contributed. Circumvallate papillae also formed in some explants in which either the geniculate or petrosal ganglia had been removed. However, placodal structures failed to mature into papillary structures even by 96 h in explants in which both ganglia had been removed. Our results demonstrate that an in vitro model using branchial arch explants supports the morphogenesis of an epithelial placode through the formation of a definite papillary structure, the circumvallate papilla, with an integrated nerve. Our results also indicate that, whereas the initial stages in gustatory papillae formation, the formation of a placode, are nerve-independent, the maturation of the placodal structure to form a papilla requires the presence of an intact nerve.     

The coexistence of TrkA with putative transmitter agents and calcium-binding proteins in the vagal and glossopharyngeal sensory neurons of the adult rat

The presence of the neurotrophin receptor, TrkA, in neurochemically identified vagal and glossopharyngeal sensory neurons of the adult rat was examined. TrkA was colocalized with calcitonin gene-related peptide (CGRP), parvalbumin, or calbindin D-28k in neurons of the nodose, petrosal and/or jugular ganglia. In contrast, no TrkA-immunoreactive (ir) neurons in these ganglia colocalized tyrosine hydroxylase-ir. About one-half of the TrkA-ir neurons in the jugular and petrosal ganglia contained CGRP-ir, whereas only a few of the numerous TrkA-ir neurons in the nodose ganglion contained CGRP-ir. Although 43% of the TrkA-ir neurons in the nodose ganglion contained calbindin D-28k-ir, few or no TrkA-ir neurons in the petrosal or jugular ganglia were also labeled for either calcium-binding protein. These data show distinct colocalizations of TrkA with specific neurochemicals in vagal and glossopharyngeal sensory neurons, and suggest that nerve growth factor (NGF), the neurotrophin ligand for TrkA, plays a role in functions of specific neurochemically defined subpopulations of mature vagal and glossopharyngeal sensory neurons.     

Coexistence of S100β and putative transmitter agents in vagal and glossopharyngeal sensory neurons of the rat

The coexistence of S100β with calcitonin gene-related peptide (CGRP), substance P (SP), somatostatin (SOM), nicotinamide adenosine dinucleotide phosphate-diaphorase (NADPH-d), and tyrosine hydroxylase (TH) was examined in the glossopharyngeal and vagal sensory ganglia. S100β immunoreactive (-ir) neurons in the jugular and petrosal ganglia frequently colocalized CGRP- or SP-ir, whereas S100β-ir neurons in the nodose ganglion infrequently contained CGRP- or SP-ir. No S100β-ir neurons in the jugular and petrosal ganglia showed SOM-ir while the small number of SOM-ir neurons in the nodose ganglion colocalized S100β-ir. Many neurons in the nodose ganglion colocalized S100β-ir and NADPH-d activity, whereas S100β-ir neurons in the jugular and nodose ganglia infrequently contained NADPH-d activity. S100β- and TH-ir were frequently colocalized in nodose ganglion but not in petrosal or jugular ganglion neurons. These findings suggest relationships between S100β and specific putative transmitters in functions of subpopulations of vagal and glossopharyngeal sensory neurons.     

Transganglionic transport of HRP from the circumvallate papilla of the rat

To learn whether horseradish peroxidase (HRP) injections in gustatory papillae on the tongue can be used to study central topographical projections of taste buds and papillae, injections were made into the circumvallate papilla in rats. Labeled central projections after papilla injections were compared to projections after applying HRP to the cut glossopharyngeal nerve. Papilla injections result in HRP transport by afferent and efferent fibers of the glossopharyngeal nerve, and the pattern of central projections is similar to that after labeling the cut nerve. Projections include a separation in the brainstem of afferent, dorsally located fibers and efferent, ventrally located fibers. Afferent fibers project to the solitary nucleus and the trigeminal system. Efferent projections label muscle motorneurons in the nucleus ambiguus and the cells of origin of parasympathetic preganglionic fibers, which from the inferior salivatory nucleus. The parasympathetic neurons labeled after papilla projections are preganglionic fibers to Remak's ganglia in the tongue; post-ganglionic fibers of these ganglia are the secretomotor supply to the von Ebner's glands. In summary, injections of HRP into gustatory papillae reliably label central projections of the papilla and can be used for studies to discern topography in central projections of the taste system. Injections into the circumvallate papilla also have demonstrated that the parasympathetic neurons innervating von Ebner's glands are located in the inferior salivatory nucleus.     

Coexistence of calbindin D-28k and NADPH-diaphorase in vagal and glossopharyngeal sensory neurons of the rat

The presence and coexistence of calbindin D-28k-immunoreactivity (ir) and nicotinamide adenosine dinucleotide phosphate (NADPH)-diaphorase activity (a marker of neurons that are presumed to convert L-arginine to L-citrulline and nitric oxide) were examined in the glossopharyngeal and vagal sensory ganglia (jugular, petrosal and nodose ganglia) of the rat. Calbindin D-28k-ir nerve cells were found in moderate and large numbers in the petrosal and nodose ganglia, respectively. Some calbindin D-28k-ir nerve cells were also observed in the jugular ganglion. NADPH-diaphorase positive nerve cells were localized to the jugular and nodose ganglia and were rare in the petrosal ganglion. A considerable portion (33–51%) of the NADPH-diaphorase positive neurons in these ganglia colocalized calbindin D-28k-ir. The presence and colocalization of calbindin D-28k-ir and NADPH-diaphorase activity in neurotransmitter-identified subpopulations of visceral sensory neurons were also studied. In all three ganglia, calcitonin gene-related peptide (CGRP)-ir was present in many NADPH-diaphorase positive neurons, a subset of which also contained calbindin D-28k-ir. In the nodose ganglion, many (42%) of tyrosine hydroxylase (TH)-ir neurons also contained NADPH diaphorase activity but did not contain calbindin D-28k-ir. These data are consistent with a potential co-operative role for calbindin D-28k and NADPH-diaphorase in the functions of a subpopulation of vagal and glossopharyngeal sensory neurons.     

Alterations in size, number, and morphology of gustatory papillae and taste buds in BDNF null mutant mice demonstrate neural dependence of developing taste organs.

Sensory ganglia that innervate taste buds and gustatory papillae (geniculate and petrosal) are reduced in volume by about 40% in mice with a targeted deletion of the gene for brain-derived neurotrophic factor (BDNF). In contrast, the trigeminal ganglion, which innervates papillae but not taste buds on the anterior tongue, is reduced by only about 18%. These specific alterations in ganglia that innervate taste organs make possible a test for roles of lingual innervation in the development of appropriate number, morphology, and spatial pattern of fungiform and circumvallate papillae and associated taste buds. We studied tongues of BDNF null mutant and wild-type littermates and made quantitative analyses of all fungiform papillae on the anterior tongue, the single circumvallate papilla on the posterior tongue, and all taste buds in both papilla types. Fungiform papillae and taste buds were reduced in number by about 60% and were substantially smaller in diameter in mutant mice 15-25 days postnatal. Remaining fungiform papillae were selectively concentrated in the tongue tip region. The circumvallate papilla was reduced in diameter and length by about 40%, and papilla morphology was disrupted. Taste bud number in the circumvallate was reduced by about 70% in mutant tongues, and the remaining taste buds were smaller than those on wild-type tongues. Our results demonstrate a selective dependence of taste organs on a full complement of appropriate innervation for normal growth and morphogenesis. Effects on papillae are not random but are more pronounced in specific lingual regions. Although the geniculate and petrosal ganglia sustain at least half of their normal complement of cell number in BDNF -/- mice, remaining ganglion cells do not substitute for lost neurons to rescue taste organs at control numbers. Whereas gustatory ganglia and the taste papillae initially form independently, our results suggest interdependence in later development because ganglia derive BDNF support from target organs and papillae require sensory innervation for morphogenesis.     

Calretinin-containing neurons which co-express parvalbumin and calbindin D-28k in the rat spinal and cranial sensory ganglia; triple immunofluorescence study

The co-expression of calretinin with parvalbumin and calbindin D-28k was examined in the rat cranial and spinal sensory ganglia by triple immunofluorescence method. In the trigeminal and nodose ganglia, 9% and 5% of calretinin-immunoreactive neurons, respectively, also contained both parvalbumin- and calbindin D-28k immunoreactivity. These neurons had large cell bodies. In the trigeminal ganglion, they were restricted to the caudal portion. Such neurons were evenly distributed throughout the nodose ganglion. The co-expression could not be detected in the dorsal root, jugular or petrosal ganglia. Nerve fibers which co-expressed all the three calcium-binding proteins were observed in the inferior alveolar nerve but not the infraorbital nerve or palate. In the periodontal ligament, these nerve fibers formed Ruffini-like endings. These findings suggest that (1) the co-expression in trigeminal neurons is intimately related to their peripheral receptive fields; (2) the three calcium-binding proteins (calretinin, parvalbumin, calbindin D-28k) co-expressed in the trigeminal neurons may have mechanoreceptive function in the periodontal ligament.     

The neural induction of taste buds in the salivary ducts of the lingual gland of von Ebner

Taste buds appear in the vallate papilla when tongue grafts are combined with sensory ganglia grafts in the anterior chamber of rats' eyes. After 50 days, many of the tongue grafts (but not the ganglion grafts) developed cysts, the papilla atrophied, and fewer taste buds were found in them than at earlier postoperative times. A study was therefore undertaken to determine whether exteriorizing the tongue graft (i.e., placing it in the confines of a corneal biopsy opening with the epithelial surface of the papilla facing outward) would better preserve the morphology of long-term grafts. In isogenic adult Lewis rats, tongue grafts were exteriorized alone or with nodose ganglia and examined after 90 to 100 days. The tongue grafts were better preserved irrespective of whether they were transplanted with or without a nodose ganglion. In addition, taste buds were now found in the epithelium of the ducts of von Ebner's lingual salivary gland. These glands lie just below the vallate papilla and because their ducts empty into the base of the papilla, portions of the ducts are unavoidably contained in tongue grafts. It appeared that intrinsic nerve fibers of the eye, just like nerve fibers from ganglia, induced the formation of ductal buds since ductal buds appeared in ten of 12 tongue grafts transplanted without added ganglia. Of significance was the observation that intense cholinesterase activity was seen in intragemmal nerve fibers of ganglionic neurons whereas no such activity was seen in intragemmal fibers when they were derived from intrinsic nerve fibers of the eye. These results indicate that exteriorization of tongue grafts better preserves their structure, that epithelium in the ducts of von Ebner's gland can be transformed into taste bud cells, that intrinsic nerve fibers of the eye can induce taste bud formation, and that there are two types of sensory neurons (based on intragemmal cholinesterase activity of their nerve fibers) which can cause taste bud development.     

Comparison of capsaicin-evoked calcium transients between rat nodose and jugular ganglion neurons

The objectives of this study were to describe the size distribution of capsaicin-sensitive neurons in nodose and jugular ganglia and to determine whether there is a difference in capsaicin sensitivity between these two types of ganglia. Functional identification was made by measurement of the capsaicin-evoked calcium (Ca2+) transients in cultured vagal sensory neurons of young adult Sprague-Dawley rats using the Fura-2-based ratiometric imaging technique. In the first study series, cells on the second day of culture were perfused with capsaicin solution (10(-7) M) for 15 s, and the Ca2+ transients were continuously recorded before, during, and after the capsaicin challenge. Out of 603 viable neurons, 57.5% were capsaicin-sensitive; the percentages of capsaicin-sensitive cells in the nodose and jugular ganglia were 59.8% and 55.4%, respectively. Capsaicin sensitivity predominated in the small- and medium-sized neurons; the capsaicin-sensitive cells generally had a diameter less than 35 microm in both types of ganglia. Although the results did not indicate any differences in the size distribution of capsaicin-sensitive neurons between the two ganglia, results of our second study series showed that a near-maximal concentration of capsaicin (3 x 10(-6) M) evoked a significantly greater peak Ca2+ transient in jugular neurons (382.5 +/- 85.5 nM) than in nodose neurons (134.3 +/- 17.5 nM). In summary, our results showed that an increase in cell diameter was accompanied by a decreasing trend in percentage of capsaicin-sensitive neurons in both vagal ganglia. Capsaicin at high concentration evoked a greater peak Ca2+ transient in jugular ganglion neurons, despite no difference in the responses to KCl between these two types of ganglion neurons.     

Co-localization of μ-opioid receptor-like immunoreactivity with substance P-LI, calcitonin gene-related peptide-LI and nitric oxide synthase-LI in vagal and glossopharyngeal afferent neurons of the rat

Co-localization of μ-opioid receptor (MOR)-like immunoreactivity (-LI) with substance P (SP)-LI, calcitonin gene-related peptide (CGRP)-LI and nitric oxide synthase (NOS)-LI in the nodose, petrosal and jugular ganglia was examined in the rat by a double immunofluorescence histochemical method. About 0.6%, 41% and 95% of neurons with MOR-LI, respectively, in the nodose, petrosal and jugular ganglia showed SP-LI; about 2%, 51% and 66% of MOR-like immunoreactive neurons displayed CGRP-LI in the nodose, petrosal and jugular ganglia, respectively. In addition, about 59% of MOR-like immunoreactive neurons in the nodose ganglia displayed NOS-LI, whereas no NOS-LI was detected in the petrosal or jugular ganglion. These data provide evidence for co-localization of MOR-LI with SP-LI, CGRP-LI and NOS-LI in the vagal and glossopharyngeal afferent neurons, and suggest that MOR may regulate the release of SP, CGRP and nitric oxide from the visceral primary afferent terminals in the nucleus of the solitary tract of the rat.     

ASIC3-immunoreactive neurons in the rat vagal and glossopharyngeal sensory ganglia

ASIC3-immunoreactivity (ir) was examined in the rat vagal and glossopharyngeal sensory ganglia. In the jugular, petrosal and nodose ganglia, 24.8%, 30.8% and 20.6% of sensory neurons, respectively, were immunoreactive for ASIC3. These neurons were observed throughout the ganglia. A double immunofluorescence method demonstrated that many ASIC3-immunoreactive (ir) neurons co-expressed calcitonin gene-related peptide (CGRP)- or vanilloid receptor subtype 1 (VRL-1)-ir in the jugular (CGRP, 77.8%; VRL-1, 28.0%) and petrosal ganglia (CGRP, 61.7%; VRL-1, 21.5%). In the nodose ganglion, however, such neurons were relatively rare (CGRP, 6.3%; VRL-1, 0.4%). ASIC3-ir neurons were mostly devoid of tyrosine hydroxylase in these ganglia. However, some ASIC3-ir neurons co-expressed calbindin D-28k in the petrosal (5.5%) and nodose ganglia (3.8%). These findings may suggest that ASIC3-containing neurons have a wide variety of sensory modalities in the vagal and glossopharyngeal sensory ganglia.     

Kv1.2-immunoreactive primary sensory neurons in the rat trigeminal ganglion

Immunohistochemistry for Kv1.2, a subunit of voltage-gated K(+) channels, was performed on the trigeminal ganglion (TG). Immunoreactivity (ir) was detected in half (48%) the TG neurons. These neurons were mostly medium-sized to large (range 137.6-2664.8 microm(2), mean+/-S.D. 892.6+/-413.3 microm(2)). A double immunofluorescence method also revealed co-expression of Kv1.2 and parvalbumin. Half (54%) the Kv1.2-immunoreactive (ir) neurons exhibited parvalbumin-ir, and parvalbumin-ir neurons mostly showed Kv1.2-ir (95%). Kv1.2-ir neurons which co-expressed CGRP-ir were rare in this ganglion. Some 40% of TG neurons retrogradely labeled from the facial skin exhibited Kv1.2-ir, whereas ir was detected in 16% of those labeled from the tooth pulp. The present study indicates that Kv1.2-ir TG neurons include low-threshold mechanoreceptors and nociceptors which innervate the facial skin and tooth pulp, respectively.