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In vivo recordings from rat geniculate ganglia: taste response properties of individual greater superficial petrosal and chorda tympani neurones.

Coding of gustatory information is complex and unique among sensory systems; information is received by multiple receptor populations located throughout the oral cavity and carried to a single central relay by four separate nerves. The geniculate ganglion is the location of the somata of two of these nerves, the greater superficial petrosal (GSP) and the chorda tympani (CT). The GSP innervates taste buds on the palate and the CT innervates taste buds on the anterior tongue. To obtain requisite taste response profiles of GSP neurones, we recorded neurophysiological responses to taste stimuli of individual geniculate ganglion neurones in vivo in the rat and compared them to those from the CT. GSP neurones had a distinct pattern of responding compared to CT neurones. For example, a small subset of GSP neurones had high response frequencies to sucrose stimulation, whereas no CT neurones had high response frequencies to sucrose. In contrast, NaCl elicited high response frequencies in a small subset of CT neurones and elicited moderate response frequencies in a relatively large proportion of GSP neurones. The robust whole-nerve response to sucrose in the GSP may be attributable to relatively few, narrowly tuned neurones, whereas the response to NaCl in the GSP may relate to proportionately more, widely tuned neurones. These results demonstrate the diversity in the initial stages of sensory coding for two separate gustatory nerves involved in the ingestion or rejection of taste solutions, and may have implications for central coding of gustatory quality and concentration as well as coding of information used in controlling energy, fluid and electrolyte homeostasis.

Action Potentials↗

Each sensory nerve arising from the geniculate ganglion expresses a unique fingerprint of neurotrophin and neurotrophin receptor genes.

Neurons in the geniculate ganglion, like those in other sensory ganglia, are dependent on neurotrophins for survival. Most geniculate ganglion neurons innervate taste buds in two regions of the tongue and two regions of the palate; the rest are cutaneous nerves to the skin of the ear. We investigated the expression of four neurotrophins, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT-3), and NT-4, and five neurotrophin receptors, trkA, trkB, trkC, p75, and truncated trkB (Trn-B) in single sensory neurons of the adult rat geniculate ganglion associated with the five innervation fields. For fungiform papillae, a glass pipette containing biotinylated dextran was placed over the target papilla and the tracer was iontophoresed into the target papilla. For the other target fields, Fluoro-Gold was microinjected. After 3 days, geniculate ganglia were harvested, sectioned, and treated histochemically (for biotinylated dextran) or immunohistochemically (for Fluoro-Gold) to reveal the neurons containing the tracer. Single labeled neurons were harvested from the slides and subjected to RNA amplification and RT-PCR to reveal the neurotrophin or neurotrophin receptor genes that were expressed. Neurons projecting from the geniculate ganglion to each of the five target fields had a unique expression profile of neurotrophin and neurotrophic receptor genes. Several individual neurons expressed more than one neurotrophin receptor or more than one neurotrophin gene. Although BDNF is significantly expressed in taste buds, its primary high affinity receptor, trkB, was not prominently expressed in the neurons. The results are consistent with the interpretation that at least some, perhaps most, of the trophic influence on the sensory neurons is derived from the neuronal somata, and the trophic effect is paracrine or autocrine, rather than target derived. The BDNF in the taste bud may also act in a paracrine or autocrine manner on the trkB expressed in taste buds, as shown by others.

Animals↗

Morphology of the nerve endings in laryngeal mucosa of the horse.

To discuss the significance of laryngeal sensation on various disorders of the horse, we studied the morphological and topographical characteristics of sensory structures in the laryngeal mucosa using immunohistochemistry and immunoelectron microscopy. Various sensory structures, i.e. glomerular endings, taste buds and intraepithelial free nerve endings, were found in the laryngeal mucosa by immunohistochemistry for protein gene product 9.5 (PGP 9.5) and neurofilament 200kD (NF200). Glomerular nerve endings were distributed mainly in the epiglottic mucosa; some endings were also found in the arytenoid region arising from thick nerve fibres running through the subepithelial connective tissue. Some terminals directly contacted the epithelial cells. Taste buds were distributed in the epithelium of the epiglottis and aryepiglottic fold. In the whole mount preparation, the taste buds were supplied by the terminal branching of the thick nerve fibres. In some cases, the taste buds were arranged around the opening of the duct of the epiglottic glands. The intraepithelial free nerve endings were found to be immunoreactive for substance P (SP) and calcitonin gene-related peptide (CGRP). These nerve endings were surrounded by the polygonal stratified epithelial cells in the supraglottic region, and by the ciliated cells in the subglottic region. The density of the intraepithelial free nerve endings was highest in the corniculate process of the arytenoid region and lowest in the vocal cord mucosa. The densities of CGRP- and SP-immunoreactive nerve endings in the arytenoid region were (mean +/- s.d.) 30.6+/-12.0 and 10.0+/-4.9 per unit epithelial length (1 mm), respectively and in the vocal fold mucosa, 1.1+/-0.9 and 0.8+/-0.7, respectively. Approximately one half of the CGRP immunoreactive nerve endings were immunoreactive for SP, and most SP-immunoreactive nerve endings were also immunoreactive for CGRP. Well-developed subepithelial plexus with numerous intraepithelial fibres were observed in flat or round mucosal projections that existed on the corniculate process of the arytenoid region. In conclusion, the laryngeal mucosa of the horse seems to have morphology- and/or location-dependent sensory mechanisms against various endo-and exogenious stimuli.

Animals↗

Development of taste organs in Rana temporaria. Transmission and scanning electron microscopic study.

In tadpoles of Rana temporaria two successive generations of taste organs: taste buds (TBs) and taste disks were studied. The TBs are located in the apices of oral premetamorphic papillae. Each TB consists of vertically elongated receptor cells (probably taste) and supporting cells. No basal (Merkel) cells were observed there. Morphologically, two types of receptor cells have been distinguished in a TB. Complete synaptic structures were observed in the TBs between the 30th and 41st of Gosner's (1960) developmental stages, though synaptic vesicles occur already at stage 26. The taste disks consist of taste cells, supporting cells and basal cells. Dense-core synaptic vesicles occur in the gustatory cells at stage 41, but complete synaptic structures were not observed earlier than at stage 45. Synaptic junctions observed at some of the gustatory cells seem to be reciprocal (bidirectional).

Animals↗

Gustducin is a taste-cell-specific G protein closely related to the transducins.

A novel G protein alpha-subunit (alpha-gustducin) has been identified and cloned from taste tissue. alpha-Gustducin messenger RNA is expressed in taste buds of all taste papillae (circumvallate, foliate and fungiform); it is not expressed in non-sensory portions of the tongue, nor is it expressed in the other tissues examined. alpha-Gustducin most closely resembles the transducins (the rod and cone photoreceptor G proteins), suggesting that gustducin's role in taste transduction is analogous to that of transducin in light transduction.

Amino Acid Sequence↗

Transgenic labeling of taste receptor cells in model fish under the control of the 5'-upstream region of medaka phospholipase C-beta 2 gene.

Vertebrate taste receptor cells express signaling molecules such as taste receptors and effectors to convert taste stimuli to inner cellular signals. Phospholipase C-beta2 (PLC-beta2) is an effector enzyme that is necessary to transduce taste signals in the mouse. It was shown that a subset of the plc-beta2 expressing cells also express taste receptor molecules, T1Rs or T2Rs, in mammals and fish. To label plc-beta2 expressing cells in the model fish species, we constructed a transgene by linking the 5'-upstream region of the medaka plc-beta2 gene to a green fluorescent protein (GFP) gene. The resulting transgenic medaka exhibited GFP signals in taste buds of the lips and the pharyngeal region. Detailed observation revealed that the GFP signals were in a subpopulation of taste bud cells, and co-localized with the transcript of endogenous plc-beta2 gene. Zebrafish introduced with the same transgene showed GFP signals in a subpopulation of taste bud cells of the lips and the pharyngeal region as in the case of medaka. This is the first report of successful labeling of taste receptor cells in two model fish species under the control of the plc-beta2 promoter. This promoter will be a useful genetic tool to study the vertebrate taste system in general.

Animals↗

Three-dimensional fine structure of the lingual papillae and their connective tissue cores in the human tongue.

The three-dimensional structure of the four types of lingual papillae and their connective tissue cores (CTCs) in the human tongue was studied by scanning electron microscopy after removal of the epithelial cell layer, and was compared to those of other mammalian species. Filiform papillae are densely distributed on the dorsal surface of the anterior two-thirds of the tongue. Each filiform papilla has numerous slender protrusions on the top. After removal of the epithelium, the CTC of the filiform papilla has a columnar primary core with 10-30 rod-shaped small secondary cores surrounding an upper central depression. A few long spine-like protrusions are frequently found in the center of this upper depression. The number, thickness and length of each secondary protrusion of the filiform CTC show some variations at the region distributed on the tongue. Fungiform papillae with spherical heads are scattered among these filiform papillae and are numerous at the anterior margin of the tongue. The CTC of the fungiform papilla shows a coralliform structure with numerous small rod-shaped protrusions on the lateral surface, and its branched top has flat areas with a few small round depressions harboring taste buds. It must be emphasized that human fungiform papilla has some taste buds not only in the young, but also in the adult age. Several to 12 vallate papillae are distributed in front of the terminal groove and had the CTC of pinecone-like structure with numerous small thorns (secondary connective tissue cores). Foliate papillae consist of 10-15 parallel folds at the posterior margin of the tongue. The CTC of the foliate papillae appears as ridges and grooves. Small protrusions are scattered on the surface of the ridges. The underside view of the exfoliated epithelium of the vallate as well as the foliate papillae revealed numerous taste buds arranged in single file in the lateral epithelium. The dorsal surface of the root of the tongue has evenly distributed short rod-shaped protrusions of the connective tissue.

Adolescent↗

A comparative study of three methods for the estimation of total plasmalogens in lingual taste epithelium and other tissues.

The total plasmalogen content of lingual and other tissues was analyzed using the iodine-addition (Method 1), the p-nitrophenylhydrazone (Method 2), and the two-dimensional thin layer chromatography procedure (Method 3). Methods 1 and 2 were simple, rapid and reproducible, yielding values usually in close agreement with each other, and values higher than those of Method 3. Method 3 exhibited poor reproducibility. All three methods were of comparable sensitivity (less than 20 nmol of total plasmalogen per sample). According to Methods 1 and 2, there was more total plasmalogen in lingual epithelium containing taste buds compared with lingual epithelium devoid of taste buds. Plasmalogen content of bovine and rat brain, heart and liver agreed with literature values.

Animals↗

Adrenergic signalling between rat taste receptor cells.

In taste buds, synaptic transmission is traditionally thought to occur from taste receptor cells to the afferent nerve. This communication reports the novel observation that taste receptor cells respond to adrenergic stimulation. Noradrenaline application inhibited outward potassium currents in a dose-dependent manner. This inhibition was mimicked by the beta agonist isoproterenol and blocked by the beta antagonist propranolol. The alpha agonists clonidine and phenylephrine both inhibited the potassium currents and elevated intracellular calcium levels. Inwardly rectifying potassium currents were unaffected by adrenergic stimulation. Experiments using the RT-PCR technique demonstrate that lingual epithelium expresses multiple alpha (alpha1a, alpha1b, alpha1c, alpha1d, alpha2a, alpha2b, alpha2c) and beta (beta1, beta2) subtypes of adrenergic receptors, and immunocytochemistry localized noradrenaline to a subset of taste receptor cells. Collectively, these data imply strongly that adrenergic transmission within the taste bud may play a paracrine role in taste physiology.

Adrenergic alpha-Agonists↗

Long-term recordings from afferent taste fibers.

The receptor cells of taste buds have a life span of about 10 days but it is not known if response characteristics of these receptors alter during the turnover cycle. To examine taste cell responses over time, a micromachined polyimide sieve electrode array was implanted between the cut ends of the rat chorda tympani nerve, which then regenerated through the electrode array. Long-term stable recordings from regenerated single afferent fibers innervating taste buds were possible using this technique for up to 21 days. Responses to taste stimuli recorded from the same fiber changed with time. The changes occurred in both the magnitude of response and the relative response profiles to four chemical stimuli, NaCl, sucrose, HCl, and quinine HCl. These changes in response characteristics were hypothesized to result from changes in the taste receptor cells as the receptor cells turnover in the taste buds.

Action Potentials↗

Taste receptor cells that discriminate between bitter stimuli.

Recent studies showing that single taste bud cells express multiple bitter taste receptors have reignited a long-standing controversy over whether single gustatory receptor cells respond selectively or broadly to tastants. We examined calcium responses of rat taste receptor cells in situ to a panel of bitter compounds to determine whether individual cells distinguish between bitter stimuli. Most bitter-responsive taste cells were activated by only one out of five compounds tested. In taste cells that responded to multiple stimuli, there were no significant associations between any two stimuli. Bitter sensation does not appear to occur through the activation of a homogeneous population of broadly tuned bitter-sensitive taste cells. Instead, different bitter stimuli may activate different subpopulations of bitter-sensitive taste cells.

Animals↗

Fine structure of the nasal barbel of the channel catfish, Ictalurus punctatus.

Ultrathin sections of the nasal barbel of the channel catfish, Ictalurus punctatus, were studied in the electron microscope and the fine structure was compared to that of barbels of other teleosts and to the mandibular (dentary) barbels of I. punctatus. While the overall histology of the nasal barbel is similar to that of barbels described previously, this study revealed far greater cellular complexity and variability than was previously reported. A layer of stratified epidermal cells rests upon a connective tissue dermis containing a cartilage rod, a large number of nerve fibers and numerous blood vessels, fibroblasts and pigment cells. Taste buds are present in the epidermal layer. This layer was found to contain probably 16 kinds of cell types, several of which may represent transitional stages, in addition to taste bud cells. Observations were made pertaining to innervation and cell types in the taste buds. A new terminology for designating the barbels of I. punctatus is suggested.

Animals↗

mRNAs for PRPs, statherin, and histatins in von Ebner's gland tissues.

A search was made for expression of genes for proline-rich proteins (PRPs) and other salivary-type proteins, including statherin and histatins, in taste-bud tissues of mice and primates because of previous genetic findings in mice (Azen et al., 1986) that Prp and taste genes for certain bitter substances are either the same or closely linked. Taste-bud tissues and other tissues were tested for specific mRNAs with labeled DNA probes by Northern blotting and in situ hybridization. It was found that PRP mRNAs were present in von Ebner's glands of mice and macaques, and that there was a much greater degree of PRP mRNA induction in mouse parotid (16-fold) than in von Ebner's gland (two-fold) after in vivo isoproterenol stimulation. This difference may be due, in part, to differences in autonomic nerve innervation. Statherin and histatin mRNAs were found in macaque taste-bud tissues containing von Ebner's gland, and statherin protein was found in human von Ebner's gland by immunohistochemistry. The finding of PRP gene expression in von Ebner's gland, whose secretions have been suggested to play a role in taste stimulation, adds further support to a possible function of PRPs in bitter tasting. The possible functions of statherin and histatins in von Ebner's gland secretions may be related to statherin's regulation of salivary calcium and histatins' antibacterial and antifungal properties.

Animals↗

Three-dimensional architecture of the subepithelial connective tissue layer in the mouse retromolar region.

The three-dimensional architecture of the subepithelial connective tissue layer in the mouse retromolar region was studied by scanning electron microscopy after removal of the epithelial layer. The connective tissue papillae (CTP) of the taste bud papillae (the retromolar papillae) appeared as a barnacle-like in form and had the round depression, corresponding to the site of taste bud, on their top. The CTP of the retromolar papillae and the openings of the salivary glands were concentrated collectively in the narrow zone just posterior to the molar teeth. This topographical relationship seems to reflect a physiological function; i.e., the food ground by the molar teeth is dissolved in saliva and stimulates the taste buds in the retromolar papillae. Further, the conical or cylindrical CTP were arranged around the CTP of the retromolar papillae. It is suggested that the conical or cylindrical CTP play an important role in increasing the epithelial-connective tissue interface and in resisting the excessive force during mastication.

Animals↗

Neural basis of developing salt taste sensation: response changes in fetal, postnatal, and adult sheep.

To learn whether salt taste responses change during mammalian development, we recorded from multifiber preparations of the chorda tympani while stimulating the anterior tongue in sheep fetuses, lambs, and adults. Stimuli were 0.5 M NH4Cl, KCl, NaCl, and LiCl, and 0.05-0.75 M concentration series of the first three salts. Ultrastructural studies were made of taste buds at different ages to determine whether morphological elements such as microvilli and tight junctions are present in young fetuses. Substantial changes occur in relative salt taste responses, throughout development. In fetuses that are beginning the last third of gestation, NaCl and LiCl elicit much smaller response magnitudes than NH4Cl and KCl. Throughout the rest of gestation and postnatally, the NaCl and LiCl responses gradually increase in magnitude relative to NH4Cl and KCl. In adults, NaCl, LiCl, and NH4Cl all elicit similar response magnitudes and KCl is less effective as a taste stimulus. At ages when response ratios for the 0.5 M salts are changing, there are no changes in shapes of the response/concentration functions for individual salts. Furthermore, microvilli are present on taste bud cell apices and tight junctions are found between cells in the youngest fetuses studied. Therefore, initial stimulus-receptor membrane contacts are probably similar to those in adults. Our data suggest that different membrane components interact with the various monochloride salts and that taste receptors contain different proportions of these various membrane components at different developmental stages. Therefore young taste bud cells do not have the same salt response characteristics as mature cells, and a changing neural substrate underlies development of salt taste function, both pre- and postnatally.

Animals↗

Expression of group II metabotropic glutamate receptors in rat gustatory papillae.

Glutamate is one candidate for the neurotransmitters and/or neuromodulators involved in taste signaling in taste buds. Group II metabotropic glutamate receptors (mGluRs: mGluR2 and mGluR3) are known to function as presynaptic receptors that regulate the release of glutamate and/or other neurotransmitters in the central nervous system. Group II mGluRs are negatively linked to adenylyl cyclase through Galphai subunits and thereby reduce the turnover of cAMP. In rat taste tissues, a subset of adenylyl-cyclase-8-expressing taste cells coexpress the Galphai subunits gustducin and Galphai2. However, the expression patterns of group II mGluRs in rat taste tissues have not yet been elucidated. We have therefore examined the expression patterns of mGluR2, mGluR3, and gustducin in rat gustatory tissues. Reverse transcription/polymerase chain reaction assays have revealed that mGluR2 and mGluR3 mRNAs are expressed in the circumvallate papillae. In situ hybridization analyses have detected positive signals for mGluR2 and mGluR3 mRNAs only in the circumvallate taste buds. Among the fungiform, foliate, and circumvallate papillae, an antibody against mGluR2/3 labels a subset of taste bud cells and nerve fibers immediately beneath the taste lingual epithelium. Double-labeling experiments have demonstrated that mGluR2/3-positive cells coexpress gustducin. These results indicate that mGluR2 and mGluR3 are coupled to Galphai subunits and play roles in glutamate-mediated signaling in taste transductions.

Animals↗

NaOH cell maceration/scanning electron microscopic studies on the connective tissue core in the incisive portion of the mouse hard palate.

NaOH cell-maceration method was applied to the oral surface of the mouse hard palate to demonstrate the three-dimensional architecture of the connective tissue core (CTC) in the incisive portion. The CTC of the incisive papilla was inverted T-shaped, and was partially fused to that of the first antemolar ruga. Between the CTC of the incisive papilla and that of the first antemolar ruga, the incisive foramen opened in the shaped of crescent hiatus. Further, the round depressions, corresponding to the site of the taste buds, were situated exclusively in the anteromedial margin of the incisive foramen and in the oral aspect of the CTC of the incisive papilla close to the foramen. Such topographical relationship between the taste buds and the incisive foramen seems to be correlated to the discriminatory function of the taste buds. In the lateral sides of the incisive papilla, the connective tissue layer formed the conspicuous serrated ridges. It is suggested that the serrated ridges are interdigitated with the furrows in the basal surface of the epithelium, and act to prevent the epithelium from slipping during mastication.

Animals↗

The role of innervation in induction and differentiation of taste organs: introduction and background.

To establish lingual receptive fields that are the basic unit of taste function, ganglion cells must extend neurites of peripheral and central targets and form connections. This symposium concerns developmental interactions between the geniculate, trigeminal and petrosal ganglia and peripheral taste organs, the gustatory papillae and resident taste buds. Investigators present data from organ and tissue culture, from mice with targeted gene deletions and from grafting experiments, in pursuit of principles that direct early innervation of the taste system. The lingual ganglia and the taste papillae initially develop independently, but then become reciprocally dependent as ganglia drive neurotrophin support from gustatory papillae and the papillae require sensory innervation for growth and morphogenesis. The issue of subsequent taste bud induction is discussed with results from amphibian and mammalian models, yielding conclusions that are not yet totally convergent. However, an essential role for sensory innervation in mammalian taste bud differentiation and acquisition of appropriate quantitative relations between ganglion cells and target organs is clearly demonstrated. A working outline is presented for periods of ganglion cell/target organ independence and interdependence during early innervation of the peripheral taste system.

Animals↗