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PubMed2005

Expression of synaptotagmin 1 in the taste buds of rat gustatory papillae.

Synapses between taste receptor cells and primary sensory afferent fibers transmit the output signal from taste buds to the central nervous system. The synaptic vesicle cycle at the synapses involves vesicle docking, priming, fusion, endocytosis, and recycling. Many kinds of synaptic vesicle proteins participate in synaptic vesicle cycles. One of these, synaptotagmin 1, binds Ca(2+) phospholipids with high affinity and plays a role in Ca(2+) regulated neurotransmitter release in the central and peripheral nervous systems. However, the expression patterns of synaptotagmin 1 in rat taste tissues have not been determined. We therefore examined the expression patterns of synaptotagmin 1 and several cell specific markers of type II and III cells in rat taste buds. RT-PCR assay showed that synaptotagmin 1 mRNA was expressed in circumvallate papillae. In fungiform, foliate, and circumvallate papillae, the antibody against synaptotagmin 1 yielded the labeling of a subset of taste bud cells and intra- and subgemmal nerve processes. Double labeled experiments showed that synaptotagmin 1 positive cells co-expressed type III cell markers, PGP 9.5, and NCAM. Intragemmal nerve processes positive for synaptotagmin 1 co-expressed PGP 9.5. Conversely, all synaptotagmin 1 expressing cells did not co-expressed type II cell markers, PLCbeta2, or gustducin. These results show that synaptotagmin 1 may play some regulatory roles in vesicle membrane fusion events with the plasma membrane at the synapses of type III cells in rat taste buds.

Animals↗
PubMed1977

Intraperitoneal transplants of taste buds in the newt.

Autografts of tongue onto the liver contained taste buds without nerves for up to 30 days. However, these denervated taste buds were generally smaller than normal and distorted in structure. Nerve fibers invaded the graft from the liver and after 30 days normal appearing taste buds were found only in implants of tongue which contained extensive nerve fibers. Thus it appears that certain visceral nerves can maintain and possibly induce taste bud formation in implants of tongue. This is interpreted as additional evidence for the trophic influence of nerves upon taste buds. Moreover, this ability to maintain taste buds is not restricted to gustatory nerves in the newt.

Animals↗
PubMed1980

Renewal of taste bud cells in rat circumvallate papillae.

The life span of taste bud cells in rat circumvallate papillae was measured by autoradiography after labeling them with a pulse of [3H]thymidine. Specimens of circumvallate papillae were taken daily 1.5-18.5 days after the isotope was administered; thereafter, specimens were taken on alternate days until 25.5 days. For each time interval, the number of labeled cell nuclei was counted in 200-450 taste buds and plotted as the ratio of labeled cells/taste bud v. time faster injection of [3H]TdR. In all, 6958 taste buds were counted. The total number of labeled cells (dark plus light) per taste bud reached peaks at 6.5, 13.5 and 20.5 days. The curve for the number of labeled dark cells/bud had essentially the same shape as that for total cells. The number of labeled light cells/bud reached a modest peak at 6.5 days and slowly declined to a plateau for the remainder of the experiment. The data show that an average of 2 days elapsed after injection before labeled dark cells entered the bud and they spent an average of 7 days in the non-proliferating taste bud compartment; thus, the life span of the dark cell was 9 days. The life span of the light cell was difficult to estimate quantitatively, but this cell type was labeled at a much slower rate than dark cells and is assumed to have a signifcantly longer tenure in the taste bud.

Animals↗
PubMed2005

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.

Animals↗
PubMed1992

Intercellular signaling in Necturus taste buds: chemical excitation of receptor cells elicits responses in basal cells.

1. Taste cells in intact taste buds in slices of Necturus lingual epithelium were impaled with microelectrodes for intracellular recording. Two types of cells were investigated: taste receptor cells and basal cells. 2. Impaling cells in the apical end of taste buds resulted in intracellular records from taste receptor cells. Applying short pulses (100- to 200-ms duration) of 140 mM KCl solution to the apical pore elicited receptor potentials in the taste receptor cells. 3. Impaling cells in the base of the taste bud resulted in intracellular records from taste receptor cells and basal cells. KCl applied to the taste pore elicited responses in the basal region that varied greatly in both magnitude and time of onset. The latency of these responses (time of onset compared with the onset of the receptor potential) ranged from 0 to hundreds of milliseconds. 4. Impaled cells were identified by injecting Lucifer yellow after recording KCl responses for 21 cells. KCl responses recorded from identified basal cells all had latencies of greater than 75 ms. KCl responses from identified receptor cells all had latencies of less than 75 ms. 5. One explanation for the long latency of KCl responses recorded in basal cells is that the responses represent postsynaptic potentials. In agreement with this interpretation, long-latency responses, but not short-latency responses, were reversibly reduced by the Ca antagonist Cd (1 mM, 10- to 20-min bath exposure). 6. Long-latency responses also differed from short-latency responses in their voltage dependence. Short-latency responses had the same voltage dependence as apically recorded receptor potentials, increasing with hyperpolarization from resting potential with an extrapolated reversal potential near 0 mV. Long-latency responses were much less dependent on voltage in this range. 7. We measured the spread of exogenously applied KCl with potassium-sensitive electrodes. Long-latency responses were not generated by diffusion of applied KCl to the basal region of the taste bud. A small transient increase in extracellular potassium occurred at the base of the taste bud after chemostimulation at the apical pore. This increase was due to depolarization-evoked release of potassium from taste cells and did not cause the long-latency responses in basal cells. 8. We conclude that short-latency (less than 75 ms) responses recorded from cells situated in the bases of taste buds are electrotonically conducted receptor potentials generated at the apical region. Long-latency (greater than 75 ms) responses are consistent with recording postsynaptic responses in basal cells.

Animals↗
PubMed1981

Fine structure of taste buds in the human fungiform papilla.

The fine structure of taste buds on fungiform papillae from man was examined with particular reference to different cell types and to cell-nerve contacts. Most of the elongated cells of the taste bud extended from the basal lamina to the apical pore, terminating in irregular microvilli. The cells contained clear apical vesicles, mitochondria, filament bundles and Golgi cisternae. Unmyelinated nerve fibers, containing mitochondria as well as both dense-cored and clear synaptic-type vesicles, were scattered throughout the whole taste bud but were most numerous at the base. The nerve fibers were either in simple appositional contact or in mesaxonal contact with the taste bud cell. Typical synapses with the postsynaptic densities in the taste bud cell were sometimes seen. In addition, single taste bud cells were observed to have more than one type of contact with nerve fibers. As in the Old World monkey Cynomolgus it was not possible to distinguish different cell types described earlier in other species.

Adult↗
PubMed2005

Morphological and biochemical heterogeneity in facial and vagal nerve innervated taste buds of the channel catfish, Ictalurus punctatus.

In catfish, the facial nerve innervates taste buds distributed over the entire body including the barbels, while the glossopharyngeal and vagal nerves innervate oropharyngeal taste buds. Facial nerve innervated taste buds (FITBs) are thought to be involved in food detection and localization, while glossopharyngeal and vagal nerve innervated taste buds (VITBs) evaluate the palatability of food prior to ingestion. Physiological studies indicate that both oral and extra-oral taste buds detect sapid substances such as amino acids and nucleotides, but the facial taste system is more sensitive to some of these substances. The anatomical, molecular, and/or physiological mechanisms underlying the functional differences in these two gustatory pathways remain to be identified. In the current investigation we compare the basic morphological features of FITBs and VITBs and the distribution of the following metabolites: gamma-aminobutyric acid (GABA), glutamate, aspartate, alanine, taurine, and glutathione. Vagal innervated taste buds are significantly longer and narrower than FITBs, with fewer taste cells and a smaller nerve plexus. Each of the metabolites examined was heterogeneously distributed in taste cells with notably more GABA positive cells present in the VITBs. Patterns of metabolite colocalization suggest the presence of several taste cell subtypes. The morphological and metabolite differences noted between FITBs and VITBs provide a potential anatomical basis for the previously noted differences in physiological sensitivity.

Amino Acids↗
PubMed1996

Embryonic taste buds develop in the absence of innervation.

It has been hypothesized that taste buds are induced by contact with developing cranial nerve fibers late in embryonic development, since descriptive studies indicate that during embryonic development taste cell differentiation occurs concomitantly with or slightly following the advent of innervation. However, experimental evidence delineating the role of innervation in taste bud development is sparse and equivocal. Using two complementary experimental approaches, we demonstrate that taste cells differentiate fully in the complete absence of innervation. When the presumptive oropharyngeal region was taken from a donor axolotl embryo, prior to its innervation and development of taste buds, and grafted ectopically on to the trunk of a host embryo, the graft developed well-differentiated taste buds. Although grafts were invaded by branches of local spinal nerves, these neurites were rarely found near ectopic taste cells. When the oropharyngeal region was raised in culture, numerous taste buds were generated in the complete absence of neural elements. Taste buds in grafts and in explants were identical to those found in situ both in terms of their morphology and their expression of calretinin and serotonin immunoreactivity. Our findings indicate that innervation is not necessary for complete differentiation of taste receptor cells. We propose that taste buds are either induced in response to signals from other tissues, such as the neural crest, or arise independently through intrinsic patterning of the local epithelium.

Ambystoma↗
PubMed1988

Human taste bud density across adult age groups.

Some of the subjective variability attributed to taste experience could be related to wide variations of taste bud density. Studies of taste perception show a direct relationship between sensation and the number of receptors. Taste bud densities are quantified in this study using light microscopy to reconstruct two regions of 18 human cadaver tongues. Specimens came from male and female cadavers representing three age groups: young adults, middle-aged adults, and older adults. The results show a range of more than 100-fold in taste bud density that is evenly distributed among age groups and sexes. The disparity is not attributable to the state of health of the adults prior to death, and it is corroborated in the literature. Differences in taste bud density that extend across age groups probably confound some inferences about the effects of aging on taste sensitivity that are derived from cross-sectional studies of human populations. It is not clear from the data whether or not human taste bud density in individuals and in populations is stable or changing with time.

Adult↗
PubMed1979

Distribution of taste buds on fungiform and circumvallate papillae of bovine tongue.

The distribution of taste buds on the fungiform and circumvallate papillae of the cow tongue has been determined. The two tongues studied were from Holstein-Friesian cows four to six years of age; they contained 14,765 and 21,691 taste buds, respectively. The tip of the tongue is well supplied with fungiform papillae, and the posterior portion contains the circumvallate papillae. The midportion of the tongue contains relatively few taste papillae. The fungiform papillae contained 1,580 and 1,838 taste buds on the two tongues, respectively, and the circumvallate papillae were estimated to contain 13,185 and 19,853 taste buds. The highest concentration of taste buds therefore occurs in the circumvallate papillae; these relatively few papillae contain approximately 90% of the taste buds. On a circumvallate papilla, taste buds are found only on the papillary sidewall, with none either on the apical surface of the papilla or on the outer wall of the moat.

Animals↗
PubMed2006

Mice lacking the p75 receptor fail to acquire a normal complement of taste buds and geniculate ganglion neurons by adulthood.

Brain-derived neurotrophic factor and neurotrophin-4 are required for normal taste bud development. Although these neurotrophins normally function via the tyrosine kinase receptor, trkB, they also bind to the pan-neurotrophin receptor, p75. The goal of the present study was to determine whether the p75 receptor is required for the development or maintenance of a full complement of adult taste buds. Mice with p75 null mutations lose 34% of their circumvallate taste buds, 36% of their fungiform papillae, and 26% of their fungiform taste buds by adulthood. The reduction of taste buds in the adult circumvallate papilla was similar to that observed previously at postnatal day 7 (Fan et al. Brain Res Dev Brain Res 2004;150:23-39). Taken together, these findings indicate that the p75 receptor is critical for the development of a full complement of taste buds, but is not required for maintenance of circumvallate taste buds in adulthood. Immunolabeling for p75 was not observed in taste buds, indicating that p75 signaling influences taste bud number indirectly. Geniculate ganglion neurons, which provides innervation to fungiform taste buds, express the p75 receptor. Mice with p75 null mutations also have fewer neurons in the geniculate ganglion. Together, these results suggest that the p75 receptor is important for the survival of geniculate neurons and geniculate neuron survival is required for the development of a full complement of taste buds by adulthood.

Aging↗
PubMed1998

A method for in-situ tight-seal recordings from single taste bud cells of mice.

In order to investigate taste transduction mechanisms, we developed a method to irrigate the receptor and basolateral membranes of mouse taste bud cells with different solutions under tight-seal recording conditions. A peeled tongue epithelium with taste bud cells was mounted on a recording platform designed to separate irrigating solutions for each membrane. The mucosal surface (receptor membrane side) of the peeled epithelium facing an inner chamber was always irrigated with deionized water or stimulating solutions, and the serosal surface (basolateral membrane side) was irrigated with a physiological saline solution. A recording electrode was placed on the basolateral membrane of a taste bud cell under an upright-microscope with a x 40-water-immersed objective. Investigated taste bud cells generated action potentials, and 1 M glucose, 200 mM NaCl, and 10 mM quinine elicited inward current. Irrigation with deionized water for more than 1 h had no effect. The resistance of the peeled tongue epithelium was 1570 +/- 343 omega cm2. These results show that the peeled tongue epithelium protects basolateral membranes from deionized water or stimulating solutions as the tongue epithelium does in situ and that this method is suitable to investigate the role of each membrane in taste transduction.

Action Potentials↗
PubMed2001

Ultrastructure of the taste buds in the blind cave fish Astyanax jordani ("Anoptichthys") and the sighted river fish Astyanax mexicanus (Teleostei, Characidae).

This study describes the ultrastructure of the taste buds of the sighted river fish Astyanax mexicanus and of the blind cave fish Astyanax jordani (= Anoptichthys) (Teleostei, Characiformes, Characidae). In Astyanax and Anoptichthys, taste buds occur in the epithelia of the lips, oral cavity, and, in Anoptichthys, lower jaw. Both possess three types of taste buds: type I (elevated), type II (slightly elevated), and type III taste buds (not elevated or sunken). The taste buds are up to 60 microm high and up to 35 microm wide. The taste bud's sensory epithelium consists of 100--130 elongated cells: light cells, dense-cored-vesicles (dcv) -cells, dark cells, and degenerating cells. The dcv-cells are rich in dense-cored vesicles and are described for the first time in a teleostean taste bud. At the taste bud's base, there lie two to three basal cells. The basal cells of type I and type II taste buds have microvillus (spine)-like processes, in contrast to those of type III taste buds. The taste bud's nerve fiber plexus is situated between the bases of the elongated taste bud cells and the basal cells. Afferent synapses occur between dcv-cells and basal cells (presynaptic sides) and axons (postsynaptic side). Indistinct synapses occur between light cells and dark cells (presynaptic sides) and axons (postsynaptic side). The nerve fiber plexes of Anoptichthys type II and type III taste buds contain significantly more axon profiles than those of Astyanax. This may be associated with a compensatory improvement of the sense of taste in the blind, cave-dwelling fish.

Animals↗
PubMed1992

Immunocytochemical survey of putative neurotransmitters in taste buds from Necturus maculosus.

To investigate synaptic mechanisms in taste buds and collect information about synaptic transmission in these sensory organs, we have examined taste buds of the mudpuppy, Necturus maculosus for the presence of neurotransmitters and neuromodulators. Immunocytochemical staining at the light microscopic level revealed the presence of serotonin-like and cholecystokinin-like (CCK) immunoreactivity in basal cells in the taste bud. Nerve fibers innervating taste buds were immunoreactive for vasoactive intestinal peptide-like (VIP), substance P-like, and calcitonin gene-related peptide-like (CGRP) or compounds closely related to these substances. Immunoreactivity for tyrosine hydroxylase (TH) and choline acetyltransferase (ChAT) in the taste cells and nerve fibers was absent. These data suggest that serotonin, CCK, VIP, substance P, and CGRP are involved in synaptic transmission or neuromodulation in the peripheral organs of taste. No evidence was found for cholinergic or adrenergic mechanisms on the basis of the absence of immunocytochemical staining for key enzymes involved in these two transmitter systems.

Animals↗
PubMed1990

Fine structure of the taste bud in guinea pigs. I. Cell characterization and innervation patterns.

Guinea pig taste buds were observed by transmission electron microscopy with special reference to cell types and innervation. The taste bud comprised four distinct cell types: basal, type I, type II, and type III cells. Basal cells, residing at the baso-lateral region of the taste bud without extending to the taste pore, were considered precursors of the other types of cells. The rest were all spindle-shaped cells reaching apically to the taste pit. Type I cells were characterized by the darkest appearance of the cytoplasm, apically possessing large, electron-dense granules and basally enveloping intragemmal nerves. This cell type, intervening between the other types of cells, was postulated to be sustentacular in nature. Type II cells, the largest and lightest cells in the taste bud, possessed a conspicuous stack of smooth endoplasmic reticulum above the nucleus. Due to their intimate and specialized relationships with nerves, the type II cells were presumed to receive an efferent innervation. Type III cells made synaptic contacts with nerves and contained dense-cored vesicles, which accumulated in the synaptic areas. This finding strongly suggests a gustatory function for the cells. The occurrence of such numerous peptidergic-type granules gathering to gustatory synapses as demonstrated in this report has not been recorded in previous papers on mammalian taste buds. The nerve terminals on the type III cell also contained synaptic vesicles, thus suggesting a reciprocal synapse here. The taste bud often included degenerating cells which were demonstrated to be phagocytosed by extrinsic cells identified as macrophages.

Animals↗
PubMed2004

[Experimental study of the fungiform papilla and taste bud regeneration following microsurgical repair of lingual nerve in rat].

PURPOSE: To investigate the changes of taste buds following injury to lingual nerve and the regeneration of the fungiform papillae and taste buds following microsurgical epineurial anastomosis of transecting injured lingual nerves in rats. METHODS: We observed the numbers and shapes of the fungiform papillae and taste buds with stereomicroscope, light microscope, and scanning electron microscope at 20 and 100 days after the clamp injury to lingual nerve, or the transecting injury to lingual nerve with/without immediate microsurgical epineurial anastomosis of the injured lingual nerve in rats. RESULTS: The fungiform papillae and taste buds degenerated, atrophied and their numbers diminished obviously at 20 days following either the clamp injury or transecting injury to the lingual nerve. The fungiform papillae and taste buds didn't regenerate spontaneously at 100 days following transection of the lingual nerve without microsurgery. The degenerated fungiform papillae and taste buds regenerated and recovered completely at 100 days following both clamp injury to the lingual nerve and transection of the lingual nerve with immediate microsurgical epineurial anastomosis. CONCLUSION: The degenerated fungiform papillae and taste buds have good ability to regenerate spontaneously following clamp injury to the lingual nerve; the degenerated fungiform papillae and taste buds can regenerate completely with immediate microsurgical epineurial anastomosis of the transected lingual nerve. The quantity and morphology of fungiform papillae and taste buds can be used as objective indicators in the function rehabilitation of injured lingual nerve.

Animals↗
PubMed1993

HVEM ultrastructural analysis of mouse fungiform taste buds, cell types, and associated synapses.

We have used high voltage electron microscopy and computer-generated three-dimensional reconstructions from serial sections to elucidate the structure of taste bud cells and their associated synapses in fungiform taste buds of the mouse. Five fungiform taste buds (two of which were serially sectioned) were examined with the high-voltage electron microscope (HVEM). We identified the synaptic connections from taste cells onto sensory nerve fibers and classified the presynaptic taste cells based on previously established ultrastructural criteria. From those data we have distinguished dark, intermediate, and light cells in murine fungiform taste buds. Synapses in murine fungiform taste buds are fewer in number, but contain many more vesicles than synapses in either foliate or circumvallate taste buds. Synapses in mouse circumvallate and foliate taste buds typically contain a few to several synaptic vesicles per section, whereas fungiform synapses may have in excess of 100 vesicles per profile. The significance of these differences in the numbers of synapses and synaptic structure between fungiform and circumvallate/foliate synapses is not known. Based on the small number of synapses observed in fungiform taste buds, we speculate that fungiform taste buds have only a few cells transducing sensory stimuli at any given time. Alternatively, communication of sensory information from the taste receptor cells to the afferent nerve fibers may be mediated by some other mechanism(s) in addition to classical chemical synapses.

Animals↗
PubMed1999

Differential expression of carbohydrate blood-group antigens on rat taste-bud cells: relation to the functional marker alpha-gustducin.

An afferent nerve fiber supplying a taste bud receives input from several taste receptor cells, yet is predominantly responsive to one of the classic taste qualities (salt, acid, sweet, or bitter). This specificity requires recognition between taste receptor cells and nerve fibers that may be mediated by surface markers correlating with function. In an effort to identify potential markers, we used immunofluorescence and confocal microscopy to examine expression of the oligosaccharide blood-group antigens Lewis(b), A, and H type 2 in taste buds of the rat oral cavity. We compared the distributions of these antigens with that of alpha-gustducin, a G-protein subunit implicated in responses to sweet- and bitter-tasting substances. The A and Lewis(b) antigens were present only on spindle-shaped cells whose apical processes reached the taste pore. These antigens were not present on epithelial cells surrounding taste buds, and Lewis(b) was not found elsewhere in the digestive tract. Lewis(b) and A were not removed by lipid extraction, suggesting that they are present on glycoproteins rather than glycolipids. All Lewis(b)-positive cells expressed alpha-gustducin, but only a fraction of alpha-gustducin-positive cells expressed Lewis(b). The fraction of taste-bud cells expressing Lewis(b) decreased in the order: vallate papillae > foliate papillae > nasoincisor duct. The epiglottis had almost no taste-bud cells that expressed Lewis(b). The A antigen appeared on taste-bud cells that also expressed alpha-gustducin in the order: foliate and vallate papillae > nasoincisor duct and epiglottis > fungiform papillae. In addition, the A antigen was present on many cells that lacked alpha-gustducin in foliate and vallate papillae. In vallate papillae, cells expressed either A or Lewis(b), but not both. Lewis(b) appears to be restricted to differentiated light cells that also express alpha-gustducin and may be involved in intercellular interactions of these cells.

Animals↗