An Airway Protection Program Revealed by Sweeping Genetic Control of Vagal Afferents.

Sensory neurons initiate defensive reflexes that ensure airway integrity. Dysfunction of laryngeal neurons is life-threatening, causing pulmonary aspiration, dysphagia, and choking, yet relevant sensory pathways remain poorly understood. Here, we discover rare throat-innervating neurons (∼100 neurons/mouse) that guard the airways against assault. We used genetic tools that broadly cover a vagal/glossopharyngeal sensory neuron atlas to map, ablate, and control specific afferent populations. Optogenetic activation of vagal P2RY1 neurons evokes a coordinated airway defense program-apnea, vocal fold adduction, swallowing, and expiratory reflexes. Ablation of vagal P2RY1 neurons eliminates protective responses to laryngeal water and acid challenge. Anatomical mapping revealed numerous laryngeal terminal types, with P2RY1 neurons forming corpuscular endings that appose laryngeal taste buds. Epithelial cells are primary airway sentinels that communicate with second-order P2RY1 neurons through ATP. These findings provide mechanistic insights into airway defense and a general molecular/genetic roadmap for internal organ sensation by the vagus nerve.

ENaC-Dependent Sodium Chloride Taste Responses in the Regenerated Rat Chorda Tympani Nerve After Lingual Gustatory Deafferentation Depend on the Taste Bud Field Reinnervated.

The chorda tympani (CT) nerve is exceptionally responsive to NaCl. Amiloride, an epithelial Na+ channel (ENaC) blocker, consistently and significantly decreases the NaCl responsiveness of the CT but not the glossopharyngeal (GL) nerve in the rat. Here, we examined whether amiloride would suppress the NaCl responsiveness of the CT when it cross-reinnervated the posterior tongue (PT). Whole-nerve electrophysiological recording was performed to investigate the response properties of the intact (CTsham), regenerated (CTr), and cross-regenerated (CT-PT) CT in male rats to NaCl mixed with and without amiloride and common taste stimuli. The intact (GLsham) and regenerated (GLr) GL were also examined. The CT responses of the CT-PT group did not differ from those of the GLr and GLsham groups, but did differ from those of the CTr and CTsham groups for some stimuli. Importantly, the responsiveness of the cross-regenerated CT to a series of NaCl concentrations was not suppressed by amiloride treatment, which significantly decreased the response to NaCl in the CTr and CTsham groups and had no effect in the GLr and GLsham groups. This suggests that the cross-regenerated CT adopts the taste response properties of the GL as opposed to those of the regenerated CT or intact CT. This work replicates the 5 decade-old findings of Oakley and importantly extends them by providing compelling evidence that the presence of functional ENaCs, essential for sodium taste recognition in regenerated taste receptor cells, depends on the reinnervated lingual region and not on the reinnervating gustatory nerve, at least in the rat.

Transnasal odontoid resection: is there an anatomic explanation for differing swallowing outcomes?

OBJECT: Swallowing dysfunction is common following transoral (TO) odontoidectomy. Preliminary experience with newer endoscopic transnasal (TN) approaches suggests that dysphagia may be reduced with this alternative. However, the reasons for this are unclear. The authors hypothesized that the TN approach results in less disruption of the pharyngeal plexus and anatomical structures associated with swallowing. The authors investigate the histological and gross surgical anatomical relationship between pharyngeal plexus innervation of the upper aerodigestive tract and the surgical approaches used (TN and TO). They also review the TN literature to evaluate swallowing outcomes following this approach. METHODS: Seven cadaveric specimens were used for histological (n = 3) and gross anatomical (n = 4) examination of the pharyngeal plexus with the TO and TN surgical approaches. Particular attention was given to identifying the location of cranial nerves (CNs) IX and X and the sympathetic chain and their contributions to the pharyngeal plexus. S100 staining was performed to assess for the presence of neural tissue in proximity to the midline, and fiber density counts were performed within 1 cm of midline. The relationship between the pharyngeal plexus, clivus, and upper cervical spine (C1-3) was defined. RESULTS: Histological analysis revealed the presence of pharyngeal plexus fibers in the midline and a significant reduction in paramedian fiber density from C-2 to the lower clivus (p < 0.001). None of these paramedian fibers, however, could be visualized with gross inspection or layer-by-layer dissection. Laterally based primary pharyngeal plexus nerves were identified by tracing their origins from CNs IX and X and the sympathetic chain at the skull base and following them to the pharyngeal musculature. In addition, the authors found 15 studies presenting 52 patients undergoing TN odontoidectomy. Of these patients, only 48 had been swallowing preoperatively. When looking only at this population, 83% (40 of 48) were swallowing by Day 3 and 92% (44 of 48) were swallowing by Day 7. CONCLUSIONS: Despite the midline approach, both TO and TN approaches may injure a portion of the pharyngeal plexus. By limiting the TN incision to above the palatal plane, the surgeon avoids the high-density neural plexus found in the oropharyngeal wall and limits injury to oropharyngeal musculature involved in swallowing. This may explain the decreased incidence of postoperative dysphagia seen in TN approaches. However, further clinical investigation is warranted.

Purinergic stimulation of carotid body efferent glossopharyngeal neurones increases intracellular Ca2+ and nitric oxide production.

The mammalian carotid body (CB) is a peripheral chemosensory organ that controls ventilation and is innervated by both afferent and efferent nerve fibres. The afferent pathway is stimulated by chemoexcitants, such as hypoxia, hypercapnia and acidosis. The efferent pathway causes inhibition of the sensory discharge via release of NO that originates mainly from neuronal nitric oxide synthase (nNOS)-positive autonomic neurones within the glossopharyngeal nerve (GPN). Recent studies in the rat indicate that these inhibitory GPN neurones and their processes express purinergic P2X receptors and can be activated by ATP, a key excitatory CB neurotransmitter. Here we tested the hypothesis that purinergic agonists stimulate a rise in [Ca(2+)]i, leading to nNOS activation and NO production in isolated GPN neurones, using the fluorescent probes fura-2 and 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate (DAF-FM DA), respectively. ATP caused a dose-dependent increase in [Ca(2+)]i in GPN neurones (EC50 ≈ 1.92 µm) that was markedly inhibited by a combination of 100 µm suramin (a non-specific P2X blocker) and 100 nm Brilliant Blue G (a selective P2X7 blocker). ATP also stimulated NO production in GPN neurones, as revealed by an increase in DAF fluorescence; this NO signal was inhibited by purinergic blockers, chelators of extracellular Ca(2+), the nNOS inhibitor l-NAME and the NO scavenger carboxy-PTIO. The P2X2/3 and P2X7 agonists α,ß,-methylene ATP and benzoyl ATP mimicked the effects of ATP. Taken together, these data indicate that ATP may contribute to negative feedback inhibition of CB sensory discharge via purinergic stimulation of NO production in efferent fibres.

Chronic hypoxia-induced acid-sensitive ion channel expression in chemoafferent neurons contributes to chemoreceptor hypersensitivity.

Previously we demonstrated that chronic hypoxia (CH) induces an inflammatory condition characterized by immune cell invasion and increased expression of inflammatory cytokines in rat carotid body. It is well established that chronic inflammatory pain induces the expression of acid-sensitive ion channels (ASIC) in primary sensory neurons, where they contribute to hyperalgesia and allodynia. The present study examines the effect of CH on ASIC expression in petrosal ganglion (PG), which contains chemoafferent neurons that innervate oxygen-sensitive type I cells in the carotid body. Five isoforms of ASIC transcript were increased ∼1.5-2.5-fold in PG following exposure of rats to 1, 3, or 7 days of hypobaric hypoxia (380 Torr). ASIC transcript was not increased in the sympathetic superior cervical ganglion (SCG). In the PG, CH also increased the expression of channel-interacting PDZ domain protein, a scaffolding protein known to enhance the surface expression and the low pH-induced current density mediated by ASIC3. Western immunoblot analysis showed that CH elevated ASIC3 protein in PG, but not in SCG or the (sensory) nodose ganglion. ASIC3 transcript was likewise elevated in PG neurons cultured in the presence of inflammatory cytokines. Increased ASIC expression was blocked in CH rats concurrently treated with the nonsteroidal anti-inflammatory drug ibuprofen (4 mg·kg(-1)·day(-1)). Electrophysiological recording of carotid sinus nerve (CSN) activity in vitro showed that the specific ASIC antagonist A-317567 (100 µM) did not significantly alter hypoxia-evoked activity in normal preparations but blocked ∼50% of the hypoxic response following CH. Likewise, a high concentration of ibuprofen, which is known to block ASIC1a, reduced hypoxia-evoked CSN activity by ∼50% in CH preparations. Our findings indicate that CH induces inflammation-dependent phenotypic adjustments in chemoafferent neurons. Following CH, ASIC are important participants in chemotransmission between type I cells and chemoafferent nerve terminals, and these proton-gated channels appear to enhance chemoreceptor sensitivity.

Antihypertensive role of glossopharyngeal nerve stimulation by nifedipine using as calcium channel blocking agent in hypertension: an experimental study.

Nifedipine is a therapeutic drug in acute attacks of hypertension because of its rapid absorption from oral mucosa. Taste receptors are innervated by glossopharyngeal nerves (GPN) as well as by facial and vagal nerves. Sensory neurons of the GPNs are localised in the petrous ganglion (PG). Transection of the taste sensitive GPN fibres causes taste bud and PG degeneration and spontaneous hypertension. In this study, the role of chemical stimulation of the taste buds of the GPN by nifedipine and its role in treatment of hypertension were investigated in rabbits. Nifedipine was dropped sublingually (20 mg) for 4 days in the study group, followed by measuring blood pressures again. Then, the lingual branches of GPNs were cut. One month later, blood pressures were measured for 4 days. All animals were sacrificed humanely at the end of the experiment, and normal and degenerated neuron densities in the petrosal ganglions were enumerated stereologically. The antihypertensive effect of nifedipine decreased after GPNs denervation, in accordance with the increase of degenerated neurons in the PG. The chemical stimulation of taste buds of the GPNs by nifedipine may be an important effect of nifedipine application in addition to its calcium channel blocking effect. The rapid decrease in blood pressure following sublingual use of nifedipine may also result from the direct stimulation of taste buds innervated by the GPNs.

Comparative study of c-Fos expression in rat dorsal vagal complex and nucleus ambiguus induced by different durations of restraint water-immersion stress.

Restraint water-immersion stress (RWIS) of rats induces vagally-mediated gastric dysfunction. The present work explored the effects of different durations of RWIS on neuronal activities of the dorsal vagal complex (DVC) and the nucleus ambiguous (NA) in rats. Male Wistar rats were exposed to RWIS for 0, 30, 60, 120, or 180 min. Then, a c-Fos immunoperoxidase technique was utilized to assess neuronal activation. Resumptively, c-Fos expression in DVC and NA peaked at 60 min of stress, subsequently decreased gradually with increasing durations of RWIS. Interestingly, the most intense c-Fos expression was observed in the dorsal motor nucleus of the vagus (DMV) during the stress, followed by NA, nucleus of solitary tract (NTS) and area postrema (AP). The peak of c-Fos expression in caudal DMV appeared at 120 min of the stress, slower than that in rostral and intermediate DMV. The c-Fos expression in intermediate and caudal NTS was significantly more intense than that in rostral NTS. These results indicate that the neuronal hyperactivity of DMV, NA, NTS and AP, the primary center that control gastric functions, especially DMV and NA, may play an important role in the disorders of gastric motility and secretion induced by RWIS.

Ionotropic glutamate receptor expression in preganglionic neurons of the rat inferior salivatory nucleus.

Glutamate receptor (GluR) subunit composition of inferior salivatory nucleus (ISN) neurons was studied by immunohistochemical staining of retrogradely labeled neurons. Preganglionic ISN neurons innervating the von Ebner or parotid salivary glands were labeled by application of a fluorescent tracer to the lingual-tonsilar branch of the glossopharyngeal nerve or the otic ganglion respectively. We used polyclonal antibodies to glutamate receptor subunits NR1, NR2A, NR2B, (NMDA receptor subunits) GluR1, GluR2, GluR3, GluR4 (AMPA receptor subunits), and GluR5-7, KA2 (kainate receptor subunits) to determine their expression in ISN neurons. The distribution of the NMDA, AMPA and kainate receptor subunits in retrogradely labeled ISN neurons innervating the von Ebner and parotid glands was qualitatively similar. The percentage of retrogradley labeled ISN neurons innervating the parotid gland expressing the GluR subunits was always greater than those innervating the von Ebner gland. For both von Ebner and parotid ISN neurons, NR2A subunit staining had the highest expression and the lowest expression of GluR subunit staining was NR2B for von Ebner ISN neurons and GluR1 for parotid ISN neurons. The percentage of NR2B and GluR4 expressing ISN neurons was significantly different between the two glands. The percentage of ISN neurons that expressed GluR receptor subunits ranged widely indicating that the distribution of GluR subunit expression differs amongst the ISN neurons. While ISN preganglionic neurons express all the GluR subunits, differences in the percentage of ISN neurons expression between neurons innervating the von Ebner and parotid glands may relate to the different functional roles of these glands.

Expression of multiple P2X receptors by glossopharyngeal neurons projecting to rat carotid body O2-chemoreceptors: role in nitric oxide-mediated efferent inhibition.

In mammals, ventilation is peripherally controlled by the carotid body (CB), which receives afferent innervation from the petrosal ganglion and efferent innervation from neurons located along the glossopharyngeal nerve (GPN). GPN neurons give rise to the "efferent inhibitory" pathway via a plexus of neuronal nitric oxide (NO) synthase-positive fibers, believed to be responsible for CB chemoreceptor inhibition via NO release. Although NO is elevated during natural CB stimulation by hypoxia, the underlying mechanisms are unclear. We hypothesized that ATP, released by rat CB chemoreceptors (type 1 cells) and/or red blood cells during hypoxia, may directly activate GPN neurons and contribute to NO-mediated inhibition. Using combined electrophysiological, molecular, and confocal immunofluorescence techniques, we detected the expression of multiple P2X receptors in GPN neurons. These receptors involve at least four different purinergic subunits: P2X2 [and the splice variant P2X2(b)], P2X3, P2X4, and P2X7. Using a novel coculture preparation of CB type I cell clusters and GPN neurons, we tested the role of P2X signaling on CB function. In cocultures, fast application of ATP, or its synthetic analog 2',3'-O-(4 benzoylbenzoyl)-ATP, caused type I cell hyperpolarization that was prevented in the presence of the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide potassium. These data suggest that ATP released during hypoxic stress from CB chemoreceptors (and/or red blood cells) will cause GPN neuron depolarization mediated by multiple P2X receptors. Activation of this pathway will lead to calcium influx and efferent inhibition of CB chemoreceptors via NO synthesis and consequent release.

ATP- and ACh-induced responses in isolated cat petrosal ganglion neurons.

Chemoreceptor (glomus) cells of the carotid body are synaptically connected to the sensory nerve endings of petrosal ganglion (PG) neurons. In response to natural stimuli, the glomus cells release transmitters, which acting on the nerve terminals of petrosal neurons increases the chemosensory afferent discharge. Among several transmitter molecules present in glomus cells, acetylcholine (ACh) and adenosine 5'-triphosphate (ATP) are considered to act as excitatory transmitter in this synapse. To test if ACh and ATP play a role as excitatory transmitters in the cat CB, we recorded the electrophysiological responses from PG neurons cultured in vitro. Under voltage clamp, ATP induces a concentration-dependent inward current that partially desensitizes during 20-30 s application pulses. The ATP-induced current has a threshold near 100 nM and saturates between 20-50 muM. ACh induces a fast, inactivating inward current, with a threshold between 10-50 muM, and saturates around 1 mM. A large part of the population of PG neurons (60%) respond to both ATP and ACh. Present results support the hypothesis that ACh and ATP act as excitatory transmitters between cat glomus cells and PG neurons.

The survival of vagal and glossopharyngeal sensory neurons is dependent upon dystonin.

The vagal and glossopharyngeal sensory ganglia and their peripheral tissues were examined in wild type and dystonia musculorum mice to assess the effect of dystonin loss of function on chemoreceptive neurons. In the mutant mouse, the number of vagal and glossopharyngeal sensory neurons was severely decreased (70% reduction) when compared with wild type littermates. The mutation also reduced the size of the circumvallate papilla (45% reduction) and the number of taste buds (89% reduction). In addition, immunohistochemical analysis demonstrated that the dystonin mutation reduced the number of PGP 9.5-, calcitonin gene-related peptide-, P2X3 receptor- and tyrosine hydroxylase-containing neurons. Their peripheral endings also decreased in the taste bud and epithelium of circumvallate papillae. These data together suggest that the survival of vagal and glossopharyngeal sensory neurons is dependent upon dystonin.

Effects of glossopharyngeal nerve section on the expression of neurotrophins and their receptors in lingual taste buds of adult mice.

The expression of neurotrophins and neurotrophin receptors is essential for the proper establishment and function of many sensory systems. To determine which neurotrophins and neurotrophin receptors are expressed in taste buds, and in taste buds of mice following denervation, antibodies directed against the neurotrophins and their receptors were applied to adult mouse gustatory tissue. Immunohistochemistry reveals that nerve growth factor (NGF)-like immunoreactive (LIR), tyrosine kinase (trk) A-LIR, trkB-LIR, and p75-LIR elongated, differentiated taste cells are present within all lingual taste buds, whereas neither neurotrophin (NT)-3- nor trkC-LIR was detected in taste cells. Double-label immunohistochemistry using markers of different taste cell types in brain-derived neurotrophic factor (BDNF)LacZ mice reveals that BDNF (beta-gal) and trkB colocalize, mainly in type III taste cells. NGF, pro-NGF, and trkA coexist in type II taste cells, i.e., those expressing phospholipase Cbeta2 (PLCbeta2). p75-LIR also is present in both BDNF and NGF taste cell populations. To determine the neural dependence of neurotrophin expression in adult taste buds, glossopharyngeal nerves were cut unilaterally. During the period of denervation (10 days to 3 weeks), taste buds largely disappear, and few neurotrophin-expressing cells are present. Three weeks after nerve transection, nerve fascicles on the operated side of the tongue exhibit BDNF-LIR, NGF-LIR, and ubiquitin carboxyl terminal hydrolase (PGP 9.5)-LIR. However, BDNF-LIR staining intensity but not NGF-LIR or PGP 9.5-LIR is increased in nerve fascicles on the operated compared with the unoperated side. Five weeks following nerve transection, NT and NT receptor expression resumes and appears normal in taste buds and nerves. These results indicate that neurotrophin expression in taste buds is dependent on gustatory innervation, but expression in nerves is not dependent on contact with taste buds.

Osteocalcin-immunoreactive neurons in the vagal and glossopharyngeal sensory ganglia of the rat.

Immunohistochemistry for osteocalcin (OC) was performed on the rat vagal and glossopharyngeal sensory ganglia. OC-immunoreactive (IR) neurons were detected in the jugular (10%), petrosal (11%) and nodose ganglia (6%). The cell size analysis demonstrated that OC-IR neurons were predominantly small to medium-sized in the jugular ganglion (mean+/-S.D.=356.3+/-192.2 microm(2), range=86.5-831.5 microm(2)). On the other hand, such neurons were medium-sized to large in the petrosal (mean+/-S.D.=725.6+/-280.7 microm(2), range=124.7-1540.4 microm(2)) and nodose ganglia (mean+/-S.D.=857.5+/-330.2 microm(2), range=367.1-1608.0 microm(2)). In the circumvallate papilla, OC-IR nerve fibers were located in the vicinity of taste buds. Some taste bud cells were also immunoreactive for the calcium-binding protein (CaBP). In the carotid body, however, OC-IR nerve fibers could not be detected. Retrograde tracing with fluorogold revealed that OC-IR nerve fibers in the circumvallate papilla mainly originated from the petrosal ganglion. These findings may suggest that OC-IR petrosal neurons have chemoreceptive function in the tongue.

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.

Motor nuclei of nerves innervating the tongue and hypoglossal musculature in a caecilian (amphibia:gymnophiona), as revealed by HRP transport.

The organization of the motor nuclei of the glossopharyngeal, vagal, occipital, first spinal and second spinal nerves of Typhlonectes natans (Amphibia: Gymnophiona: Caeciliaidae: Typhlonectinae) was studied by using horseradish peroxidase reaction staining. Each nucleus has discrete patterns of cytoarchitecture and of topography. Nuclei are elongate and some overlap anteroposteriorly. The brainstem is elongate, with no distinct demarcation of brainstem from spinal cord. The occipital nerve emerges through a separate foramen from that for the vagus and glossopharyngeal nerves in the species studied, is distinct from both, and its nucleus is more similar to spinal nuclei in cytoarchitecture. The occipital nerve fuses with spinal nerves 1 and 2 to contribute to the hypoglossal trunk. A spinal accessory nerve is absent.

Distribution of transient receptor potential channels in the rat carotid chemosensory pathway.

Glomus cells in the carotid body respond to decreases in oxygen tension of the blood and transmit this sensory information in the carotid sinus nerve to the brain via neurons in the petrosal ganglion. G-protein-coupled membrane receptors linked to phospholipase C may play an important role in this response through the activation of the cation channels formed by the transient receptor potential (TRP) proteins. In the present study, expression of TRPC proteins in the rat carotid body and petrosal ganglion was examined using immunohistochemical techniques. TRPC3, TRPC4, TRPC5, TRPC6, and TRPC7 were present in neurons throughout the ganglion. TRPC1 was expressed in only 28% of petrosal neurons, and of this population, 45% were tyrosine hydroxylase (TH)-positive, accounting for essentially all the TH-expressing neurons in the ganglion. Because TH-positive neurons project to the carotid body, this result suggests that TRPC1 is selectively associated with the chemosensory pathway. Confocal images through the carotid body showed that TRPC1/3/4/5/6 proteins localize to the carotid sinus nerve fibers, some of which were immunoreactive to an anti-neurofilament (NF) antibody cocktail. TRPC1 and TRPC3 were present in both NF-positive and NF-negative fibers, whereas TPRC4, TRPC5, and TRPC6 expression was primarily localized to NF-negative fibers. Only TRPC1 and TRPC4 were localized in the afferent nerve terminals that encircle individual glomus cells. TRPC7 was not expressed in sensory fibers. All the TRPC proteins studied were present in type I glomus cells. Although their role as receptor-activated cation channels in the chemosensory pathway is yet to be established, the presence of TRPC channels in glomus cells and sensory nerves of the carotid body suggests a role in facilitating and/or sustaining the hypoxic response.

Expression of the neural cell adhesion molecule (NCAM) and polysialic acid during taste bud degeneration and regeneration.

Taste receptor cells are replaced throughout life, accompanied by continuing synaptogenesis between newly formed taste cells and first-order gustatory fibers. The neural cell adhesion molecule (NCAM) is expressed by a subset of taste cells in adult rodents and appears on gustatory nerve fibers during development prior to differentiation of the taste buds. We employed antibodies against the extracellular domain of the NCAM polypeptide (mAb 3F4) and against polysialic acid (PSA) residues found on embryonic forms of NCAM (mAb 5A5) to investigate the relationship between the expression of these molecules and the innervation of taste buds in adult rats. In unoperated rats, anti-NCAM recognized a subset of cells within the vallate taste buds and also the fibers of the glossopharyngeal (IXth) nerve, including those innervating the gustatory epithelium. Taste bud cells did not express PSA but mAb 5A5 immunoreactivity was observed on some fibers of the IXth nerve, including a few that entered the taste buds. Bilateral crush of the IXth nerve resulted in the loss of NCAM expression from the gustatory epithelium within 8 days. As IXth nerve fibers reinnervated the epithelium, NCAM expression was seen first in the nerve, followed by increased expression in the epithelium as the taste cells differentiated from their precursors. PSA expression by fibers of the IXth nerve did not return to normal until well after the regeneration of the vallate taste buds. The present results demonstrate that taste cell expression of NCAM is dependent upon innervation by the IXth nerve and that NCAM expression appears in the nerve prior to its expression in the differentiating epithelium during regeneration.(ABSTRACT TRUNCATED AT 250 WORDS)

Substance P in the vagal sensory ganglia: localization in cell bodies and pericellular arborizations.

The distribution of Substance P-like immunoreactivity in the jugular and nodose ganglia of rabbits and pigeons has been studied using immunocytochemical staining techniques. Substance P-like immunoreactivity is localized to neuronal cell bodies and processes in the jugular and nodose ganglia, and to pericellular fiber plexi in the nodose ganglia of both species. The numbers and sizes of cells which exhibited Substance P-like immunoreactivity in each ganglion were determined using quantitative morphometric techniques. The distribution of Substance P-like immunoreactivity in the rabbit and pigeon vagal sensory ganglia is characterized by several general features. In most of the ganglia, immunoreactive neurons factor into discrete types which can be distinguished from one another, and from non-immunoreactive neurons, by size. In addition, immunoreactive nodose and jugular ganglion cells, respectively, are distinguishable on the basis of size. Finally, a considerably high percentage of immunoreactive neurons is found in the jugular ganglion than in the nodose ganglion. Substance P-like immunoreactivity was also seen in pericellular fiber plexi which encircle individual neurons in the nodose ganglion of rabbits and pigeons. These plexi are composed of varicose fibers which appear to terminate as boutons on the surface of the cells which they encircle. The distribution of Substance P-like immunoreactivity within the vagal sensory ganglia is discussed with respect to the possible peripheral targets and functions of Substance P-containing vagal afferents. Our findings suggest that Substance P-containing vagal sensory neurons are involved in a variety of visceral and somatic afferent functions.

Locations of androgen-concentrating cells in the brain of Xenopus laevis: autoradiography with 3H-dihydrotestosterone.

The distribution of hormone-concentrating cells in the brains of South African clawed frogs, Xenopus laevis, was examined autoradiographically after the administration of 3H-dihydrotestosterone. Hormone-accumulating cells were found in cranial nerve nucleus IX-X and adjacent smaller cells, a presumed medullary vestibular nucleus, a presumed sensory nucleus of cranial nerve V, dorsal tegmental area of the medulla, laminar nucleus of the torus semicircularis, ventral thalamus, and anterior pituitary. The pattern of dihydrotestosterone-labelled cells differs from previously reported results following testosterone or estradiol administration. Unlike these latter hormones, dihydrotestosterone does not accumulate in anterior preoptic or ventral infundibular nuclei. Both androgens but not estradiol label medullary motor neurons; limbic telencephalic nuclei appear to concentrate only estradiol. Hormone-concentrating brain nuclei in X. laevis have been implicated in neuro-endocrine regulation and the control of male and female reproductive behaviors.

Immunohistochemical localization of putative neurotransmitters within the feline nucleus tractus solitarii.

With the aid of immunohistochemical techniques the distribution of substance P, met-enkephalin, serotonin, somatostatin, alpha-melanocyte stimulating hormone, neurotensin and neurophysin immunoreactivities were mapped throughout the rostro-caudal extent of the cat's nucleus tractus solitarii. Three of the putative neurotransmitters (substance P, enkephalin and serotonin) were found to be widely distributed as varicose fibers and punctate structures. The densities of their immunoreactivities were plotted in a range from very dense, dense, moderate, occasional, to none, at different levels of the nucleus of the solitary tract. Substance P immunoreactivity was the most varied and dense of all the neurotransmitters studied. Its accumulations ranged from very dense in the lateral, dense in portions of the parvocellular and lateral, moderate in medial and commissural and occasional in ventrolateral and portions of the parvocellular subdivisions. Both the enkephalin and serotonin immunoreactivities had patterns similar to that of substance P immunoreactivity, although their amounts were not as great. Following colchicine treatment neurons containing substance P and enkephalin immunoreactivity were found in many subdivisions of the nucleus of the solitary tract. Somatostatin, alpha-melanocyte stimulating hormone, neurotensin and neurophysin immunoreactivities were present in the nucleus of the solitary tract as isolated varicose fibers scattered throughout the nucleus. Immunoreactive neurons were not found for these putative neurotransmitters after colchicine treatment. The presence of substance P immunoreactivity within subdivisions which receive visceral afferent input is discussed in relation to the role of substance P as a possible transmitter of the afferent limb of the vagus nerve. The distribution of enkephalin and serotonin immunoreactivities in the nucleus of the solitary tract reflect their involvement in the regulation or modulation of cardiovascular and respiratory functions. While the significance of somatostatin, alpha-melanocyte stimulating hormone, neurotensin and neurophysin immunoreactivities within the nucleus of the solitary tract is not understood at present, these substances might possibly play a role in visceral functions.