RESUMEN
KEY POINTS: Adenosine and dopamine (DA) are neuromodulators in the carotid body (CB) chemoafferent pathway, but their mechanisms of action are incompletely understood. Using functional co-cultures of rat CB chemoreceptor (type I) cells and sensory petrosal neurons (PNs), we show that adenosine enhanced a hyperpolarization-activated cation current Ih in chemosensory PNs via A2a receptors, whereas DA had the opposite effect via D2 receptors. Adenosine caused a depolarizing shift in the Ih activation curve and increased firing frequency, whereas DA caused a hyperpolarizing shift in the curve and decreased firing frequency. Acute hypoxia and isohydric hypercapnia depolarized type I cells concomitant with increased excitation of adjacent PNs; the A2a receptor blocker SCH58261 inhibited both type I and PN responses during hypoxia, but only the PN response during isohydric hypercapnia. We propose that adenosine and DA control firing frequency in chemosensory PNs via their opposing actions on Ih . ABSTRACT: Adenosine and dopamine (DA) act as neurotransmitters or neuromodulators at the carotid body (CB) chemosensory synapse, but their mechanisms of action are not fully understood. Using a functional co-culture model of rat CB chemoreceptor (type I) cell clusters and juxtaposed afferent petrosal neurons (PNs), we tested the hypothesis that adenosine and DA act postsynaptically to modulate a hyperpolarization-activated, cyclic nucleotide-gated (HCN) cation current (Ih ). In whole-cell recordings from hypoxia-responsive PNs, cAMP mimetics enhanced Ih whereas the HCN blocker ZD7288 (2 µm) reversibly inhibited Ih . Adenosine caused a potentiation of Ih (EC50 â¼ 35 nm) that was sensitive to the A2a blocker SCH58261 (5 nm), and an â¼16 mV depolarizing shift in V½ for voltage dependence of Ih activation. By contrast, DA (10 µm) caused an inhibition of Ih that was sensitive to the D2 blocker sulpiride (1-10 µm), and an â¼11 mV hyperpolarizing shift in V½ . Sulpiride potentiated Ih in neurons adjacent to, but not distant from, type I cell clusters. DA also decreased PN action potential frequency whereas adenosine had the opposite effect. During simultaneous paired recordings, SCH58261 inhibited both the presynaptic hypoxia-induced receptor potential in type I cells and the postsynaptic PN response. By contrast, SCH58261 inhibited only the postsynaptic PN response induced by isohydric hypercapnia. Confocal immunofluorescence confirmed the localization of HCN4 subunits in tyrosine hydroxylase-positive chemoafferent neurons in tissue sections of rat petrosal ganglia. These data suggest that adenosine and DA, acting through A2a and D2 receptors respectively, regulate PN excitability via their opposing actions on Ih .
Asunto(s)
Adenosina/farmacología , Dopamina/farmacología , Neuronas/efectos de los fármacos , Animales , Células Cultivadas , Técnicas de Cocultivo , Ganglio Geniculado/citología , Ganglio Geniculado/metabolismo , Canales Regulados por Nucleótidos Cíclicos Activados por Hiperpolarización/metabolismo , Neuronas/fisiología , Canales de Potasio/metabolismo , Ratas WistarRESUMEN
Afferent chorda tympani (CT) fibers innervating anterior tongue fungiform papillae have neuron cell bodies in the geniculate ganglion (GG). To characterize electrophysiological and receptive field properties, we recorded extracellular responses from single GG neurons to lingual application with chemical, thermal and mechanical stimuli. Receptive field size was mapped by electrical stimulation of individual fungiform papillae. Responses of GG neurons to room temperature chemical stimuli representing five taste qualities, and distilled water at 4⯰C and mechanical stimulation were used. Based on responses to these stimuli, GG neurons were divided into CHEMICAL, CHEMICAL/THERMAL, THERMAL and TACTILE groups. Neurons in the CHEMICAL group responded to taste stimuli but not to either cold water or stroking stimuli. CHEMICAL/THERMAL neurons responded to both taste stimuli and cold water. THERMAL neurons responded only to cold water and TACTILE neurons responded only to light stroking stimuli. The receptive field sizes for CHEMICAL, and CHEMICAL/THERMAL neurons averaged five papillae exceeding the field size of THERMAL and TACTILE neurons which averaged about two papillae. Detailed analysis of the receptive field of CHEMICAL/THERMAL neurons revealed that within one field only a subset of the fungiform papillae making up the receptive field responded to the cold stimuli, whereas the other papillae responded only to chemical stimuli. These finding demonstrate that fungiform papilla are complex sensory organs with a multisensory function suggesting a unique role in detecting and sampling food components prior to ingestion.
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Potenciales de Acción/fisiología , Ganglio Geniculado/citología , Células Receptoras Sensoriales/clasificación , Células Receptoras Sensoriales/fisiología , Lengua/inervación , Animales , Biofisica , Femenino , Fibras Nerviosas/fisiología , Técnicas de Placa-Clamp , Estimulación Física/efectos adversos , Ratas , Ratas Sprague-Dawley , Estimulación Química , Temperatura , Tacto/fisiologíaRESUMEN
Taste buds are innervated by neurons whose cell bodies reside in cranial sensory ganglia. Studies on the functional properties and connectivity of these neurons are hindered by the lack of markers to define their molecular identities and classes. The mouse geniculate ganglion contains chemosensory neurons innervating lingual and palatal taste buds and somatosensory neurons innervating the pinna. Here, we report single cell RNA sequencing of geniculate ganglion neurons. Using unbiased transcriptome analyses, we show a pronounced separation between two major clusters which, by anterograde labeling, correspond to gustatory and somatosensory neurons. Among the gustatory neurons, three subclusters are present, each with its own complement of transcription factors and neurotransmitter response profiles. The smallest subcluster expresses both gustatory- and mechanosensory-related genes, suggesting a novel type of sensory neuron. We identify several markers to help dissect the functional distinctions among gustatory neurons and address questions regarding target interactions and taste coding.Characterization of gustatory neural pathways has suffered due to a lack of molecular markers. Here, the authors report single cell RNA sequencing and unbiased transcriptome analyses to reveal major distinctions between gustatory and somatosensory neurons and subclusters of gustatory neurons with unique molecular and functional profiles.
Asunto(s)
Ganglio Geniculado/metabolismo , Neurotransmisores/metabolismo , Células Receptoras Sensoriales/metabolismo , Transcriptoma , Animales , Pabellón Auricular/inervación , Ganglio Geniculado/citología , Proteínas de Homeodominio/genética , Ratones , Proteínas del Tejido Nervioso/genética , Receptores Purinérgicos P2X2/genética , Receptores Purinérgicos P2X3/genética , Análisis de Secuencia de ARN , Análisis de la Célula Individual , ATPasa Intercambiadora de Sodio-Potasio/genética , Proteína 25 Asociada a Sinaptosomas/genética , Gusto , Papilas Gustativas , Lengua/inervación , Tacto , Factores de Transcripción/genéticaRESUMEN
Many basic characteristics of gustatory neurons remain unknown, partly due to the absence of specific markers. Some neurons in the geniculate ganglion project to taste regions in the oral cavity, whereas others innervate the outer ear. We hypothesized that the transcription factor Phox2b would identify oral cavity-projecting neurons in the geniculate ganglion. To test this possibility, we characterized mice in which Phox2b-Cre mediated gene recombination labeled neurons with tdTomato. Nerve labeling revealed that all taste neurons projecting through the chorda tympani (27%) and greater superficial petrosal nerves (15%) expressed Phox2b during development, whereas non-oral somatosensory neurons (58%) in the geniculate ganglion did not. We found tdTomato-positive innervation within all taste buds. Most (57%) of the fungiform papillae had labeled innervation only in taste buds, whereas 43% of the fungiform papillae also had additional labeled innervation to the papilla epithelium. Chorda tympani nerve transection eliminated all labeled innervation to taste buds, but most of the additional innervation in the fungiform papillae remained. Some of these additional fibers also expressed tyrosine hydroxylase, suggesting a sympathetic origin. Consistent with this, both sympathetic and parasympathetic fibers innervating blood vessels and salivary glands contained tdTomato labeling. Phox2b-tdTomato labels nerve fascicles in the tongue of the developing embryo and demonstrates a similar stereotyped branching pattern DiI-labeling.
Asunto(s)
Ganglio Geniculado/citología , Proteínas de Homeodominio/metabolismo , Células Receptoras Sensoriales/metabolismo , Factores de Transcripción/metabolismo , Animales , Toxina del Cólera/metabolismo , Nervio de la Cuerda del Tímpano/citología , Embrión de Mamíferos , Fluoresceína/metabolismo , Regulación de la Expresión Génica , Proteínas de Homeodominio/genética , Insulina/metabolismo , Proteínas con Homeodominio LIM/metabolismo , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Ratones , Ratones Transgénicos , Boca/inervación , Gusto/fisiología , Papilas Gustativas/fisiología , Lengua/inervación , Factores de Transcripción/genética , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
The vestibular and geniculate ganglia of the ear in experimental animals carry both of the tyrosine hydroxylase (TH)-positive sympathetic neurons and the neuronal nitric oxide synthase (nNOS)-positive parasympathetic neurons. With an aid of immunohistochemistry, we examined these ganglia as well as the horizontal part of the facial nerve using specimens from 10 formalin-fixed elderly cadavers. The submandibular ganglion from the same cadavers was used for the positive control for both markers. Although there was a nonspecific reaction in nuclei for the present antibody of nNOS, these ganglia were unlikely to contain either nNOS- or TH-positive neurons. However, we did not deny a possibility that the absence was a result of degeneration with aging. In contrast, the facial nerve horizontal part consistently contained both of TH-positive- and nNOS-positive fibers. These fibers might regulate blood supply to the facial nerve and the dysregulation leads to edema to elevate pressure on the nerve within its osseous canal.
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Ganglios Parasimpáticos/citología , Ganglios Simpáticos/citología , Ganglio Geniculado/citología , Neuronas/citología , Vestíbulo del Laberinto/citología , Anciano , Anciano de 80 o más Años , Cadáver , Humanos , Inmunohistoquímica , Masculino , Fibras NerviosasRESUMEN
Afferent chorda tympani (CT) fibers innervating taste and somatosensory receptors in fungiform papillae have neuron cell bodies in the geniculate ganglion (GG). The GG/CT fibers branch in the tongue to innervate taste buds in several fungiform papillae. To investigate receptive field characteristics of GG/CT neurons, we recorded extracellular responses from GG cells to application of chemical and thermal stimuli. Receptive field size was mapped by electrical stimulation of individual fungiform papillae. Response latency to electrical stimulation was used to determine fiber conduction velocity. Responses of GG neurons to lingual application of stimuli representing four taste qualities, and water at 4°C, were used to classify neuron response properties. Neurons classified as SALT, responding only to NaCl and NH4Cl, had a mean receptive field size of six papillae. Neurons classified as OTHER responded to salts and other chemical stimuli and had smaller mean receptive fields of four papillae. Neurons that responded to salts and cold stimuli, classified as SALT/THERMAL, and neurons responding to salts, other chemical stimuli and cold, classified as OTHER/THERMAL, had mean receptive field sizes of six and five papillae, respectively. Neurons responding only to cold stimuli, categorized as THERMAL, had receptive fields of one to two papillae located at the tongue tip. Based on conduction velocity most of the neurons were classified as C fibers. Neurons with large receptive fields had higher conduction velocities than neurons with small receptive fields. These results demonstrate that GG neurons can be distinguished by receptive field size, response properties and afferent fiber conduction velocity derived from convergent input of multiple taste organs.
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Nervio de la Cuerda del Tímpano/fisiología , Ganglio Geniculado/citología , Conducción Nerviosa/fisiología , Neuronas/fisiología , Células Receptoras Sensoriales/fisiología , Potenciales de Acción/fisiología , Cloruro de Amonio/farmacología , Análisis de Varianza , Animales , Estimulación Eléctrica , Femenino , Conducción Nerviosa/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Tiempo de Reacción , Células Receptoras Sensoriales/efectos de los fármacos , Cloruro de Sodio/farmacología , Estimulación Química , Gusto/fisiología , Papilas Gustativas/fisiologíaRESUMEN
The mammalian taste system is responsible for sensing and responding to the five basic taste qualities: sweet, sour, bitter, salty and umami. Previously, we showed that each taste is detected by dedicated taste receptor cells (TRCs) on the tongue and palate epithelium. To understand how TRCs transmit information to higher neural centres, we examined the tuning properties of large ensembles of neurons in the first neural station of the gustatory system. Here, we generated and characterized a collection of transgenic mice expressing a genetically encoded calcium indicator in central and peripheral neurons, and used a gradient refractive index microendoscope combined with high-resolution two-photon microscopy to image taste responses from ganglion neurons buried deep at the base of the brain. Our results reveal fine selectivity in the taste preference of ganglion neurons; demonstrate a strong match between TRCs in the tongue and the principal neural afferents relaying taste information to the brain; and expose the highly specific transfer of taste information between taste cells and the central nervous system.
Asunto(s)
Ganglio Geniculado/citología , Neuronas/fisiología , Percepción del Gusto/fisiología , Gusto/fisiología , Lengua/fisiología , Animales , Calcio/metabolismo , Ratones , Ratones Transgénicos , Papilas Gustativas/citología , Papilas Gustativas/fisiología , Lengua/citología , Lengua/inervaciónRESUMEN
HYPOTHESIS: Pretreatment with antiherpetic medications and steroids decreases likelihood of development of delayed facial paralysis (DFP) after otologic surgery. BACKGROUND: Heat-induced reactivation of herpes simplex virus type 1 (HSV1) in geniculate ganglion neurons (GGNs) is thought to cause of DFP after otologic surgery. Antiherpetic medications and dexamethasone are used to treat DFP. Pretreatment with these medications has been proposed to prevent development of DFP. METHODS: Rat GGN cultures were latently infected with HSV1 expressing a lytic protein-GFP chimera. Cultures were divided into pretreatment groups receiving acyclovir (ACV), acyclovir-plus-dexamethasone (ACV + DEX), dexamethasone alone (DEX), or untreated media (control). After pretreatment, all cultures were heated 43°C for 2 hours. Cultures were monitored daily for reactivation with fluorescent microscopy. Viral titers were determined from culture media. RESULTS: Heating cultures to 43°C for 2 hours leads to HSV1 reactivation and production of infectious virus particles (59 ± 6.8%); heating cultures to 41°C showed a more variable frequency of reactivation (60 ± 40%), compared with baseline rates of 14.4 ± 5%. Cultures pretreated with ACV showed lower reactivation rates (ACV = 3.7%, ACV + DEX = 1.04%) compared with 44% for DEX alone. Viral titers were lowest for cultures treated with ACV or ACV + DEX. CONCLUSION: GGN cultures harboring latent HSV1 infection reactivate when exposed to increased temperatures that can occur during otologic surgery. Pretreatment with ACV before heat provides prophylaxis against heat-induced HSV reactivation, whereas DEX alone is associated with higher viral reactivation rates. This study provides evidence supporting the use of prophylactic antivirals for otologic surgeries associated with high rates of DFP.
Asunto(s)
Aciclovir/uso terapéutico , Antiinflamatorios/uso terapéutico , Antivirales/uso terapéutico , Dexametasona/uso terapéutico , Parálisis Facial/etiología , Parálisis Facial/prevención & control , Procedimientos Quirúrgicos Otológicos/efectos adversos , Complicaciones Posoperatorias/prevención & control , Animales , Células Cultivadas , Ganglio Geniculado/citología , Calor , Neuronas/efectos de los fármacos , Reacción en Cadena de la Polimerasa , Ratas , Ratas Sprague-Dawley , Carga Viral , Activación Viral/efectos de los fármacos , Latencia del VirusRESUMEN
We analyzed the spike discharge patterns of two types of neurons in the rodent peripheral gustatory system, Na specialists (NS) and acid generalists (AG) to lingual stimulation with NaCl, acetic acid, and mixtures of the two stimuli. Previous computational investigations found that both spike rate and spike timing contribute to taste quality coding. These studies used commonly accepted computational methods, but they do not provide a consistent statistical evaluation of spike trains. In this paper, we adopted a new computational framework that treated each spike train as an individual data point for computing summary statistics such as mean and variance in the spike train space. We found that these statistical summaries properly characterized the firing patterns (e. g. template and variability) and quantified the differences between NS and AG neurons. The same framework was also used to assess the discrimination performance of NS and AG neurons and to remove spontaneous background activity or "noise" from the spike train responses. The results indicated that the new metric system provided the desired decoding performance and noise-removal improved stimulus classification accuracy, especially of neurons with high spontaneous rates. In summary, this new method naturally conducts statistical analysis and neural decoding under one consistent framework, and the results demonstrated that individual peripheral-gustatory neurons generate a unique and reliable firing pattern during sensory stimulation and that this pattern can be reliably decoded.
Asunto(s)
Biología Computacional/métodos , Ganglio Geniculado/citología , Neuronas/citología , Estadística como Asunto , Algoritmos , Animales , Potenciales Evocados/efectos de los fármacos , Ganglio Geniculado/efectos de los fármacos , Ganglio Geniculado/fisiología , Masculino , Neuronas/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Percepción del Gusto/efectos de los fármacos , Factores de TiempoRESUMEN
The geniculate ganglion, which provides innervation to taste buds in the anterior tongue and palate, is unique among sensory ganglia in that its neurons depend on both neurotrophin-4 (NT4) and brain-derived neurotrophic factor (BDNF) for survival. Whereas BDNF is additionally implicated in taste axon guidance at targeting stages, much less is known about the guidance role of NT4 during targeting, or about either neurotrophin during initial pathfinding. NT4 and BDNF have distinct expression patterns in vivo, raising the possibility of distinct roles. We characterized the influence of NT4 and BDNF on geniculate neurites in collagen I gels at early embryonic through postnatal stages. During early pathfinding to the tongue (embryonic days 12-13; E12-13), NT4 and BDNF promote significantly longer outgrowth than during intralingual targeting (E15-18). NT4 is more potent than BDNF at stimulating neurite outgrowth and both factors exhibit concentration optima, i.e. intermediate concentrations (0.25 ng/ml NT4 or 25 ng/ml BDNF) promote maximal neurite extension and high concentrations (10 ng/ml NT4 or 200 ng/ml BDNF) suppress it. Only partial suppression was seen at E12 (when axons first emerge from the ganglion in vivo) and postnatally, but nearly complete suppression occurred from E13 to E18. We show that cell death is not responsible for suppression. Although blocking the p75 receptor reduces outgrowth at the optimum concentrations of NT4 and BDNF, it did not reduce suppression of outgrowth. We also report that NT4, like BDNF, can act as a chemoattractant for geniculate neurites, and that the tropic influence is strongest during intralingual targeting (E15-18). NT4 does not appear to act as an attractant in vivo, but it may prevent premature invasion of the epithelium by suppressing axon growth.
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Factor Neurotrófico Derivado del Encéfalo/farmacología , Ganglio Geniculado/crecimiento & desarrollo , Factores de Crecimiento Nervioso/farmacología , Neuritas/efectos de los fármacos , Animales , Axones/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Ganglio Geniculado/citología , Ganglio Geniculado/efectos de los fármacos , Embarazo , Ratas , Receptor de Factor de Crecimiento Nervioso/biosíntesis , Receptor de Factor de Crecimiento Nervioso/genética , Lengua/embriología , Lengua/metabolismoRESUMEN
The number of neurons in the geniculate ganglion that are available to innervate taste buds is regulated by neurotrophin-4 (NT-4) and brain-derived neurotrophic factor (BDNF). Our goal for the current study was to examine the timing and mechanism of NT-4-mediated regulation of geniculate neuron number during development. We discovered that NT-4 mutant mice lose 33% of their geniculate neuronal cells between E10.5 and E11.5. By E11.5, geniculate axons have just reached the tongue and do not yet innervate their gustatory targets; thus, NT-4 does not function as a target-derived growth factor. At E11.5, no difference was observed in proliferating cells or the rate at which cells exit the cell cycle between NT-4 mutant and wild type ganglia. Instead, there was an increase in TUNEL-labeling, indicating an increase in cell death in Ntf4(-/-) mice compared with wild types. However, activated caspase-3, which is up-regulated in the absence of BDNF, was not increased. This finding indicates that cell death initiated by NT-4-removal occurs through a different cell death pathway than BDNF-removal. We observed no additional postnatal loss of taste buds or neurons in Ntf4(-/-) mice. Thus, during early embryonic development, NT-4 produced in the ganglion and along the projection pathway inhibits cell death through an activated caspase-3 independent mechanism. Therefore, compared to BDNF, NT-4 plays distinct roles in gustatory development; differences include timing, source of neurotrophin, and mechanism of action.
Asunto(s)
Factor Neurotrófico Derivado del Encéfalo/fisiología , Ganglio Geniculado/embriología , Factores de Crecimiento Nervioso/fisiología , Animales , Factor Neurotrófico Derivado del Encéfalo/genética , Caspasa 3/fisiología , Diferenciación Celular , Movimiento Celular , Supervivencia Celular , Ganglio Geniculado/citología , Ganglio Geniculado/fisiología , Ratones , Factores de Crecimiento Nervioso/genética , Neuronas/citología , Neuronas/fisiología , Papilas Gustativas/fisiologíaRESUMEN
HYPOTHESIS: Reactivation of herpes simplex virus type 1 (HSV-1) in geniculate ganglion neurons (GGNs) is an etiologic mechanism of Bell's palsy (BP) and delayed facial palsy (DFP) after otologic surgery. BACKGROUND: Several clinical studies, including temporal bone studies, antibody, titers, and intraoperative studies, suggest that reactivation of HSV-1 from latently infected GGNs may lead to both BP and DFP. However, it is difficult to study these processes in humans or live animals. METHODS: Primary cultures of GGNs were latently infected with Patton strain HSV-1 expressing a green fluorescent protein-late lytic gene chimera. Four days later, these cultures were treated with trichostatin A (TSA), a known chemical reactivator of HSV-1 in other neurons. Cultures were monitored daily by fluorescent microscopy. Titers of media from lytic, latent, and latent/TSA treated GGN cultures were obtained using plaque assays on Vero cells. RNA was harvested from latently infected GGN cultures and examined for the presence of viral transcripts using reverse transcription-polymerase chain reaction. RESULTS: Latently infected GGN cultures displayed latency-associated transcripts only, whereas lytically infected and reactivated latent cultures produced other viral transcripts, as well. The GGN cultures displayed a reactivation rate of 65% after treatment with TSA. Media from latently infected cultures contained no detectable infectious HSV-1, whereas infectious virus was observed in both lytically and latently infected/TSA-treated culture media. CONCLUSION: We have shown that cultured GGNs can be latently infected with HSV-1, and HSV-1 in these latently infected neurons can be reactivated using TSA, yielding infectious virus. These results have implications for the cause of both BP and DFP.
Asunto(s)
Parálisis Facial/etiología , Parálisis Facial/virología , Herpesvirus Humano 1 , Animales , Parálisis de Bell/etiología , Parálisis de Bell/prevención & control , Parálisis de Bell/virología , Células Cultivadas , Chlorocebus aethiops , Medios de Cultivo , Parálisis Facial/prevención & control , Ganglio Geniculado/citología , Ganglio Geniculado/virología , Proteínas Fluorescentes Verdes , Microscopía Fluorescente , Neuronas/virología , ARN Viral/biosíntesis , Ratas , Ratas Sprague-Dawley , Reacción en Cadena en Tiempo Real de la Polimerasa , Transcripción Genética , Células Vero , Activación Viral , Latencia del Virus/genéticaRESUMEN
Polycystic kidney disease 1-like 3 (Pkd1l3) is expressed specifically in sour-sensing type III taste cells that have synaptic contacts with afferent nerve fibers in circumvallate (CvP) and foliate papillae (FoP) located in the posterior region of the tongue, although not in fungiform papillae (FuP) or the palate. To visualize the gustatory neural pathways that originate from type III taste cells in CvP and FoP, we established transgenic mouse lines that express the transneuronal tracer wheat germ agglutinin (WGA) under the control of the mouse Pkd1l3 gene promoter/enhancer. The WGA transgene was accurately expressed in Pkd1l3-expressing type III taste cells in CvP and FoP. Punctate WGA protein signals appeared to be detected specifically in type III taste cells but not in other types of taste cells. WGA protein was transferred primarily to a subset of neurons located in close proximity to the glossopharyngeal (GL) nerve bundles in the nodose/petrosal ganglion (NPG). WGA signals were also observed in a small population of neurons in the geniculate ganglion (GG). This result demonstrates the anatomical connection between taste receptor cells (TRCs) in the FoP and the chorda tympani (CT) nerves. WGA protein was further conveyed to neurons in a rostro-central subdivision of the nucleus of the solitary tract (NST). These findings demonstrate that the approximately 10 kb 5'-flanking region of the mouse Pkd1l3 gene functions as a type III taste cell-specific promoter/enhancer. In addition, experiments using the pkd1l3-WGA transgenic mice reveal a sour gustatory pathway that originates from TRCs in the posterior region of the tongue.
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Canales Catiónicos TRPP/biosíntesis , Papilas Gustativas/citología , Papilas Gustativas/metabolismo , Gusto/genética , Lengua/citología , Lengua/fisiología , Animales , Canales de Calcio , Regulación del Desarrollo de la Expresión Génica , Ganglio Geniculado/química , Ganglio Geniculado/citología , Ganglio Geniculado/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Vías Nerviosas/química , Vías Nerviosas/citología , Vías Nerviosas/fisiología , Ganglio Nudoso/química , Ganglio Nudoso/citología , Ganglio Nudoso/fisiología , Canales Catiónicos TRPP/química , Canales Catiónicos TRPP/genética , Papilas Gustativas/química , Lengua/química , Aglutininas del Germen de Trigo/biosíntesis , Aglutininas del Germen de Trigo/química , Aglutininas del Germen de Trigo/genéticaRESUMEN
Geniculate ganglion (GG) cell bodies of chorda tympani (CT), greater superficial petrosal (GSP), and posterior auricular (PA) nerves transmit orofacial sensory information to the rostral nucleus of the solitary tract. We have used whole cell recording to investigate the characteristics of the Na(+) channels in isolated Fluorogold-labeled GG neurons that innervate different peripheral receptive fields. GG neurons expressed two classes of Na(+) channels, TTX sensitive (TTX-S) and TTX resistant (TTX-R). The majority of GG neurons expressed TTX-R currents of different amplitudes. TTX-R currents were relatively small in 60% of the neurons but were large in 12% of the sampled population. In a further 28% of the neurons, TTX completely abolished all Na(+) currents. Application of TTX completely inhibited action potential generation in all CT and PA neurons but had little effect on the generation of action potentials in 40% of GSP neurons. Most CT, GSP, and PA neurons stained positively with IB(4), and 27% of the GSP neurons were capsaicin sensitive. The majority of IB(4)-positive GSP neurons with large TTX-R Na(+) currents responded to capsaicin, whereas IB(4)-positive GSP neurons with small TTX-R Na(+) currents were capsaicin insensitive. These data demonstrate the heterogeneity of GG neurons and indicate the existence of a subset of GSP neurons sensitive to capsaicin, usually associated with nociceptors. Since there are no reports of nociceptors in the GSP receptive field, the role of these capsaicin-sensitive neurons is not clear.
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Potenciales de Acción/fisiología , Ganglio Geniculado/citología , Neuronas/fisiología , Canales de Sodio/metabolismo , Potenciales de Acción/efectos de los fármacos , Análisis de Varianza , Animales , Animales Recién Nacidos , Biofisica , Capsaicina/farmacología , Estimulación Eléctrica , Lectinas/metabolismo , Masculino , Neuronas/clasificación , Neuronas/efectos de los fármacos , Técnicas de Placa-Clamp , Ratas , Ratas Sprague-Dawley , Fármacos del Sistema Sensorial/farmacología , Bloqueadores de los Canales de Sodio/farmacología , Tetrodotoxina/farmacologíaRESUMEN
Geniculate ganglion (GG) cell bodies of chorda tympani (CT), greater superficial petrosal (GSP), and posterior auricular (PA) nerves transmit orofacial sensory information to the rostral nucleus of the solitary tract (rNST). We used whole cell recording to study the characteristics of the Ca(2+) channels in isolated Fluorogold-labeled GG neurons that innervate different peripheral receptive fields. PA neurons were significantly larger than CT and GSP neurons, and CT neurons could be further subdivided based on soma diameter. Although all GG neurons possess both low voltage-activated (LVA) "T-type" and high voltage-activated (HVA) Ca(2+) currents, CT, GSP, and PA neurons have distinctly different Ca(2+) current expression patterns. Of GG neurons that express T-type currents, the CT and GSP neurons had moderate and PA neurons had larger amplitude T-type currents. HVA Ca(2+) currents in the GG neurons were separated into several groups using specific Ca(2+) channel blockers. Sequential applications of L, N, and P/Q-type channel antagonists inhibited portions of Ca(2+) current in all CT, GSP, and PA neurons to a different extent in each neuron group. No difference was observed in the percentage of L- and N-type Ca(2+) currents reduced by the antagonists in CT, GSP, and PA neurons. Action potentials in GG neurons are followed by a Ca(2+) current initiated after depolarization (ADP) that may influence intrinsic firing patterns. These results show that based on Ca(2+) channel expression the GG contains a heterogeneous population of sensory neurons possibly related to the type of sensory information they relay to the rNST.
Asunto(s)
Canales de Calcio Tipo T/fisiología , Ganglio Geniculado/fisiología , Células Receptoras Sensoriales/fisiología , Potenciales de Acción/fisiología , Animales , Nervio de la Cuerda del Tímpano/citología , Nervio de la Cuerda del Tímpano/fisiología , Pabellón Auricular/inervación , Ganglio Geniculado/citología , Masculino , Potenciales de la Membrana/fisiología , Modelos Animales , Técnicas de Placa-Clamp , Ratas , Ratas Sprague-DawleyRESUMEN
Geniculate axons are initially guided to discrete epithelial placodes in the lingual and palatal epithelium that subsequently differentiate into taste buds. In vivo approaches show that brain-derived neurotrophic factor (BDNF) mRNA is concentrated in these placodes, that BDNF is necessary for targeting taste afferents to these placodes, and that BDNF misexpression disrupts guidance. We used an in vitro approach to determine whether BDNF may act directly on geniculate axons as a trophic factor and as an attractant, and whether there is a critical period for responsiveness to BDNF. We show that BDNF promotes neurite outgrowth from geniculate ganglion explants dissected from embryonic day (E) 15, E18, infant, and adult rats cultured in collagen gels, and that there is a concentration optimum for neurite extension. Gradients of BDNF derived from slow-release beads caused the greatest bias in neurite outgrowth at E15, when axons approach the immature gustatory papillae. Further, neurites advanced faster toward the BDNF bead than away from it, even if the average amount of neurotrophic factor encountered was the same. We also found that neurites that contact BDNF beads did not advance beyond them. At E18, when axons would be penetrating pregustatory epithelium in vivo, BDNF continued to exert a tropic effect on geniculate neurites. However, at postnatal and adult stages, the influence of BDNF was predominantly trophic. Our data support a role for BDNF acting as an attractant for geniculate axons during a critical period that encompasses initial targeting but not at later stages.
Asunto(s)
Factor Neurotrófico Derivado del Encéfalo/farmacología , Factores Quimiotácticos/farmacología , Ganglio Geniculado , Neuritas , Animales , Factor Neurotrófico Derivado del Encéfalo/genética , Factores Quimiotácticos/genética , Epitelio/embriología , Epitelio/crecimiento & desarrollo , Ganglio Geniculado/citología , Ganglio Geniculado/efectos de los fármacos , Ganglio Geniculado/fisiología , Neuritas/efectos de los fármacos , Neuritas/metabolismo , Neuritas/ultraestructura , Ratas , Papilas Gustativas/citología , Papilas Gustativas/embriología , Papilas Gustativas/crecimiento & desarrollo , Técnicas de Cultivo de Tejidos , Lengua/citología , Lengua/embriología , Lengua/crecimiento & desarrollo , Lengua/inervaciónRESUMEN
Neurons of the geniculate ganglion innervate taste buds located in two spatially distinct targets, the tongue and palate. About 50% of these neurons die in Bdnf(-/-) mice and Ntf4/5(-/-) mice. Bdnf(-/-)/Ntf4/5(-/-) double mutants lose 90-95% of geniculate ganglion neurons. To determine whether different subpopulations are differentially influenced by neurotrophins, we quantified neurons from two ganglion subpopulations separately and remaining taste buds at birth within each target field in wild-type, Bdnf(-/-), Ntf4/5(-/-), and Bdnf(-/-)/Ntf4/5(-/-) mice. In wild-type mice the same number of neurons innervated the anterior tongue and soft palate and each target contained the same number of taste buds. Compared to wild-type mice, Bdnf(-/-) mice showed a 50% reduction in geniculate neurons innervating the tongue and a 28% loss in neurons innervating the soft palate. Ntf4/5(-/-) mice lost 58% of the neurons innervating the tongue and 41% of the neurons innervating the soft palate. Taste bud loss was not as profound in the NT-4 null mice compared to BDNF-null mice. Tongues of Bdnf(-/-)/Ntf4/5(-/-) mice were innervated by 0 to 4 gustatory neurons and contained 3 to 16 taste buds at birth, indicating that some taste buds remain even when all innervation is lost. Thus, gustatory neurons are equally dependent on BDNF and NT-4 expression for survival, regardless of what peripheral target they innervate. However, taste buds are more sensitive to BDNF than NT-4 removal.
Asunto(s)
Factor Neurotrófico Derivado del Encéfalo/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Neuronas Aferentes/fisiología , Hueso Paladar/inervación , Gusto/fisiología , Lengua/inervación , Animales , Factor Neurotrófico Derivado del Encéfalo/genética , Ganglio Geniculado/citología , Ganglio Geniculado/embriología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Factores de Crecimiento Nervioso/genética , Neuronas/citología , Neuronas/fisiología , Hueso Paladar/embriología , Papilas Gustativas/citología , Papilas Gustativas/fisiología , Lengua/embriologíaRESUMEN
The purpose of this study was to investigate the role of response latency in discrimination of chemical stimuli by geniculate ganglion neurons in the rat. Accordingly, we recorded single-cell 5-s responses from geniculate ganglion neurons (n = 47) simultaneously with stimulus-evoked summated potentials (electrogustogram; EGG) from the anterior tongue to signal when the stimulus contacted the lingual epithelium. Artificial saliva served as the rinse solution and solvent for all stimuli [(0.5 M sucrose, 0.03-0.5 M NaCl, 0.01 M citric acid, and 0.02 M quinine hydrochloride (QHCl)], 0.1 M KCl as well as for 0.1 M NaCl +1 µM benzamil. Cluster analysis separated neurons into four groups (sucrose specialists, NaCl specialists, NaCl/QHCl generalists and acid generalists). Artificial saliva elevated spontaneous firing rate and response frequency of all neurons. As a rule, geniculate ganglion neurons responded with the highest frequency and shortest latency to their best stimulus with acid generalist the only exception. For specialist neurons and NaCl/QHCl generalists, the average response latency to the best stimulus was two to four times shorter than the latency to secondary stimuli. For NaCl-specialist neurons, response frequency increased and response latency decreased systematically with increasing NaCl concentration; benzamil significantly decreased NaCl response frequency and increased response latency. Acid-generalist neurons had the highest spontaneous firing rate and were the only group that responded consistently to citric acid and KCl. For many acid generalists, a citric-acid-evoked inhibition preceded robust excitation. We conclude that response latency may be an informative coding signal for peripheral chemosensory neurons.
Asunto(s)
Ganglio Geniculado/fisiología , Nervio Lingual/fisiología , Tiempo de Reacción/fisiología , Células Receptoras Sensoriales/fisiología , Gusto/fisiología , Animales , Ácido Cítrico/farmacología , Ganglio Geniculado/citología , Masculino , Modelos Animales , Cloruro de Potasio/farmacología , Quinina/farmacología , Ratas , Ratas Sprague-Dawley , Células Receptoras Sensoriales/clasificación , Cloruro de Sodio/farmacología , Estimulación Química , Sacarosa/farmacología , Gusto/efectos de los fármacosRESUMEN
A study was made on 18 embryos of developmental stages 13-15 (5(th) week). Serial sections made in horizontal, frontal, and sagittal planes were stained with routine histological methods and some of them were treated with silver. In embryos of stage 13, the otic vesicle is at the rhombomere 5, and close to the vesicle is the facial-vestibulocochlear ganglionic complex in which the geniculate, vestibular, and cochlear ganglion may be discerned. These ganglia are well demarcated in embryos of stage 14. In the last investigated stage (15(th)) the nerve fibres of the ganglia reach the common afferent tract.