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Location and variation in number of taste buds in human fungiform papillae.

Serial sections of 182 fungiform papillae, obtained at autopsy from 22 individuals aged 2 days to 90 years, were examined by light microscopy with regard to location and number of taste buds. The taste buds were always found on the convex, dorsal surface of the papillae but otherwise failed to display any preferential location pattern. A total of 262 taste buds, an average of 1.4 per papilla, were found. However, there was considerable variation in the occurrence of taste buds, both from papilla to papilla and from case to case. While the number of taste buds in a single papilla varied from 0 to 27, 63% of the papillae had no taste buds at all, 26% had 1-3 buds and the remainder 4 or more buds. The mean number of taste buds per papilla varied from 0 to 9 between individuals; no dependence upon sex or age could be demonstrated for this variation. The significance of these anatomical findings with regard to physiological studies on taste involving the fungiform papillae is discussed.

Adolescent↗

Uptake and release of neurotransmitter candidates, [3H]serotonin, [3H]glutamate, and [3H]gamma-aminobutyric acid, in taste buds of the mudpuppy, Necturus maculosus.

Neurotransmitters in vertebrate taste buds have not yet been identified with confidence. Serotonin, glutamate, and gamma-aminobutyric acid (GABA) have been postulated, but the evidence is incomplete. We undertook an autoradiographic study of [3H]serotonin, [3H]glutamate, and [3H]GABA uptake in lingual epithelium from the amphibian, Necturus maculosus, to determine whether taste bud cells would accumulate and release these substances. Lingual epithelium containing taste buds was incubated in low concentrations (0.4-6 microM) of these tritiated transmitter candidates and the tissue was processed for light microscopic autoradiography. Merkel-like basal taste cells accumulated [3H]serotonin. When the tissue was treated with 40 mM K+ after incubating the tissue in [3H]serotonin, cells released the radiolabelled transmitter. Furthermore, depolarization (KCl)-induced release of [3H]serotonin was Ca-dependent: if Ca2+ was reduced to 0.4 mM and 20 mM Mg2+ added to the high K+ bathing solution, Merkel-like basal cells did not release [3H]serotonin. In contrast, [3H]glutamate was taken up by several cell types, including non-sensory epithelial cells, Schwann cells, and some taste bud cells. [3H]glutamate was not released by depolarizing the tissue with 40 mM K+. [3H]GABA uptake was also widespread, but did not occur in taste bud cells. [3H]GABA accumulated in non-sensory epithelial cells and Schwann cells. These data support the hypothesis that serotonin is a neurotransmitter or neuromodulator released by Merkel-like basal cells in Necturus taste buds. The data do not support (nor rule out) a neurotransmitter role for glutamate or GABA in taste buds.

Animals↗

Taste buds: development and evolution.

The gustatory system in vertebrates comprises peripheral receptors (taste buds), innervated by three cranial nerves (VII, IX, and X), and a series of central neural centers and pathways. All vertebrates, with the exception of hagfishes, have taste buds. These receptors vary morphologically in different vertebrates but usually consist of at least four types of cells (dark, light, basal, and stem cells). An out-group analysis indicates that taste buds were restricted to the oropharynx, primitively, and that external taste buds, distributed over the head and, in some cases, even the trunk, evolved a number of times independently. The sensory neurons of the cranial nerves that innervate taste buds are believed to arise from epibranchial placodes, which are induced by pharyngeal endoderm, but it has never been demonstrated experimentally that these sensory neurons do, in fact, arise from these placodes. Although many details of the development of the innervation of taste buds are still unknown, it is now clear that taste buds are induced from either ecto- or endodermal epithelia, rather than arising from either placodes or neural crest. At present, there are two developmental models of taste bud induction: The neural induction model claims that peripheral nerve fibers induce taste buds, whereas the early specification model claims that oropharyngeal epithelium is specified by or during gastrulation and that taste buds arise from cell-cell interactions within the specified epithelium. There is now substantial evidence that the early specification model best describes the induction of taste buds.

Animals↗

Electron microscopic observation of vallate taste buds of zinc-deficient rats with taste disturbance.

Henkin et al. (1975) reported that there is a loss of the dense substance in the taste pore region, a loss of microvilli and a decrease of dark granules and microvacuolation in the taste pore buds of patients who complain of taste disturbance. In our experiment, we prepared artificially zinc-deficient rats by feeding a zinc-deficient diet and observed the same results in the taste buds of zinc-deficient rats with taste disturbance.

Animals↗

Quantitative anatomical study of taste buds in fungiform papillae of young and old Fischer rats.

To determine if differences in neural taste responses relate to taste bud loss in old age, taste buds were counted in fungiform papillae of Fischer 344 rats aged 4 to 6 months, 20 to 24 months, and 30 to 37 months. Papillae anterior to the intermolar eminence on one half of the tongue were examined in serial sections. Presence or absence of a taste bud was noted and taste bud diameter was measured. Average percentages of papillae that contained a taste bud in the three groups were 99.6, 99.3, and 94.7%. This is a significant age-related difference but actual number of taste buds lost in the oldest rats was small. Taste bud diameter did not differ with age and general anatomical characteristics of buds were similar in all groups. Thus, anatomical observations on taste bud maintenance in rats over a wide age range, coupled with neurophysiological data, demonstrate that the integrity of the peripheral gustatory system is not altered greatly in old age.

Aging↗

The distribution and origin of keratin 20-containing taste buds in rat and human.

Sections of tissues containing lingual and extra-lingual taste buds were evaluated with monoclonal antibodies against cytokeratins. In the caudal third of the rat's tongue, keratin 20 immunoreactivity was restricted to taste buds, whereas keratins 7, 8, 18, and 19 were expressed in vallate and foliate taste buds and in cells of salivary ducts that merge with these taste epithelia. Hence, antibodies against keratin 20 most clearly distinguished differentiated taste cells from all other cells. In rat epiglottis, taste buds and isolated bipolar cells were keratin-20-positive. In rat nasopalatine papilla and palate, antibodies against keratin 20 identified Merkel cells, none of which was near to the keratin-20-negative taste buds. Nor were Merkel cells present at epiglottal taste buds or the keratin-20-negative fungiform taste buds or elsewhere in rat tongue. Hence, Merkel cells make no contribution to rat fungiform, epiglottal, nasopalatine, or palatal taste buds. Human and rat keratin-20-positive tissues are reported to be endodermal derivatives with the exception of Merkel cells and luminal urothelial cells. In rats the distribution of keratin-20-positive taste buds was in full agreement with the classical view that the posterior third of the tongue is derived from endoderm (keratin-20-positive taste buds), whereas the anterior two-thirds of the tongue is derived from stomadeal ectoderm (keratin-20-negative taste buds). The equally intense keratin 20 immunoreactivity of human fungiform and vallate taste buds violates this traditional rostro-caudal segregation and suggests that endodermally derived tissues may be present in the tip of the human tongue.

Adult↗

Development and maturation of taste buds of the palatal epithelium of the rat: histological and immunohistochemical study.

Palatal taste buds are intriguing partners in the mediation of taste behavior and their spatial distribution is functionally important for suckling behavior, especially in the neonatal life. Their prenatal development has not been previously elucidated in the rat, and the onset of their maturation remains rather controversial. We delineated the development and frequency distribution of the taste buds as well as the immunohistochemical expression of alpha-gustducin, a G protein closely related to the transduction of taste stimuli, in the nasoincisor papilla (NIP) and soft palate (SP) from the embryonic day 17 (E17) till the postnatal day 70 (PN70). The main findings in the present study were the development of a substantial number of taste pores in the SP of fetal rats (60.3 +/- 1.7 out of 122.8 +/- 5.5; mean +/- SD/animal at E19) and NIP of neonatal rats (9.8 +/- 1.0 out of 44.8 +/- 2.2 at PN4). alpha-gustducin-like immunoreactivity (-LI) was not expressed in the pored taste buds of either prenatal or newborn rats. The earliest expression of alpha-gustducin-LI was demonstrated at PN1 in the SP (1.5 +/- 0.5 cells/taste bud; mean +/- SD) and at PN4 in the NIP (1.4 +/- 0.5). By age the total counts of pored taste buds continuously increased and their morphological features became quite discernible. They became pear in shape, characterized by distinct pores, long subporal space, and longitudinally oriented cells. Around the second week, a remarkable transient decrease in the total number of taste buds was recorded in the oral epithelium of NIP and SP, which might be correlated with the changes of ingestive behaviors. The total counts of cells showing alpha-gustducin-LI per taste bud gradually increased till the end of our investigation (14.1 +/- 2.7 in NIP and 12.4 +/- 2.5 in SP at PN70). We conclude that substantial development of taste buds began prenatally in the SP, whereas most developed entirely postnatal in the NIP. The present study provides evidence that the existence of a taste pore which is considered an important criterion for the morphological maturation of taste buds is not enough for the onset of the taste transduction, which necessitates also mature taste cells. Moreover, the earlier maturation of palatal taste buds compared with the contiguous populations in the oral cavity evokes an evidence of their significant role in the transmission of gustatory information, especially in the early life of rat.

Animals↗

Neural cell adhesion molecule, neuron-specific enolase and calcitonin gene-related peptide immunoreactivity in hamster taste buds after chorda tympani/lingual nerve denervation.

Hamster fungiform papilla taste buds persist in an atrophic form following sensory denervation. While atrophic and innervated taste buds are morphologically similar, it is not known whether their gemmal cells have similar molecular characteristics. Three neurochemicals, neural cell adhesion molecule, neuron-specific enolase, and calcitonin gene-related peptide have been implicated in trophic phenomena, synaptogenesis and cell recognition in neurons and sensory neuroepithelia. The present study uses immunocytochemical localization of these molecular markers to characterize normal and denervated fungiform taste buds following unilateral chorda tympani/lingual nerve denervation in hamsters. In normal taste buds, immunoreactivity to neural cell adhesion molecule, neuron-specific enolase, and calcitonin gene-related peptide was present in a group of cells located centrally in the bud as well as in fungiform nerve fibres and endings. After denervation, gemmal cell immunoreactivity to all three markers was reduced and often confined to a single or a few bud cell(s). Also, fibre staining was absent except for sparse calcitonin gene-related peptide-immunoreactive fibres associated with blood vessels and within the fungiform papillae. These remaining fibres may be autonomic or somatomotor in origin. These results indicate that sensory denervation of hamster taste buds reduces, but does not wholly eliminate the immunoreactivity of surviving gemmal cells to neural cell adhesion molecule, neuron-specific enolase, and calcitonin gene-related peptide. While the number of taste bud cells expressing the markers appears to be nerve-dependent, immunoreactivity in sensory-denervated bud cells of hamster may reflect the influence of local tissue factors.

Animals↗

The role of substance P and calcitonin gene-related peptide containing nerve fibers in maintaining fungiform taste buds in the rat after a chronic chorda tympani nerve injury.

Taste buds in the anterior part of the tongue of adult rats were denervated by unilateral resection of the chorda tympani nerve in the middle ear. Three months later one group of animals was perfused and their tongues were processed for demonstration of substance P (SP) and calcitonin gene-related peptide (CGRP) immunoreactivity. Fungiform taste buds found on the denervated side showed increased numbers of intragemmal SP- and CGRP-immunoreactive (IR) fibers compared to the normal side. Compared to the normal side, the number of taste buds appeared to be fewer on the denervated side. Moreover, taste buds on this side seemed to be only partially restored. Another group of animals was given the neurotoxin capsaicin which causes a depletion of SP and CGRP from sensory axons. The animals were perfused 2 or 3 weeks after the capsaicin treatment, and their tongues prepared for SP and CGRP immunohistochemistry or for histological examination of taste buds. Very few SP- and CGRP-IR fibers were present in capsaicin-treated animals. In these animals almost all fungiform taste buds and papillae on the chorda tympani-injured side disappeared. In contrast, normal numbers of taste buds were still present on the contralateral side where the chorda tympani innervation remained intact. It is conceivable that taste buds on the chorda tympani-innervated part of the tongue, deprived of the normal chorda tympani-innervation, can regenerate and become reinnervated by SP- and CGRP-containing fibers, and that these are essential for partially restoring and maintaining the structure of the denervated taste buds and the fungiform papillae.

Animals↗

Ecto-calcium-dependent ATPase activity of mammalian taste bud cells.

Histochemistry was utilized to characterize Ca-ATPases associated with lingual taste buds in the golden hamster. Taste buds showed elevated staining for magnesium- or calcium-dependent ATPase (Ca-ATPase) relative to the surrounding epithelium. At low calcium concentrations (0.1-0.5 mM), intracellular staining predominated. Most of the studies were conducted at calcium concentrations of > or = 10 mM, in which most of the staining was localized to the external face of plasma membranes of taste bud cells (including receptor and basal cells) located in the core of fungiform taste buds, or the entire vallate or foliate taste buds. The peripheral fungiform taste bud cells stained much less intensely, but the peripheral cells adjacent to the core showed intermediate levels. GTP and ITP were just as effective substrates as ATP. Millimolar concentrations of magnesium were as effective as calcium. Inhibitors of intracellular ATPases, including quercetin, sodium azide, and 2,4-dinitrophenol, had no effect on the staining. Therefore, the Ca-ATPase staining of plasma membranes at mM concentrations of calcium is thought to correspond to one or more ecto-Ca-ATPase activities with unknown functions. Roles related to increased energy requirements or to the possible function of ATP as a neurotransmitter or -modulator are proposed.

2,4-Dinitrophenol↗

Voltage-gated channels involved in taste responses and characterizing taste bud cells in mouse soft palates.

Taste bud cells (TBCs) on soft palates differ from those on tongues in innervation and chemosensitivity. We investigated voltage-gated channels involved in the taste responses of TBCs on mouse soft palates under in-situ tight-seal voltage/current-clamp conditions. Under the cell-attached mode, TBCs spontaneously fired action currents, which were blocked by application of 1 microM TTX to TBC basolateral membranes. Firing frequencies increased in response to taste substances applied to TBC receptor membranes. Under the whole-cell clamp mode, as expected, TBCs produced various voltage-gated currents such as TTX-sensitive Na+ currents (INa), outward currents (Iout) including TEA-sensitive and insensitive currents, inward rectifier K+ currents (Iir), and Ca2+ currents including T-type, P/Q-type, and L-type Ca2+ currents. We classified TBCs into three types based on the magnitude of their voltage-gated Na+ currents and membrane capacitance. HEX type (60% of TBCs examined) was significantly larger in Na+ current magnitude and smaller in membrane capacitance than LEX type (23%). NEX type (17%) had no Na+ currents. HEX type was equally distributed within single taste buds, while LEX type was centrally distributed, and NEX type was peripherally distributed. There were correlations between these electrophysiological cell types and morphological cell types determined by three-dimensional reconstruction. The present results show that soft palate taste buds contain TBCs with different electrophysiological properties, and suggest that their co-operation is required in taste transduction.

Action Potentials↗

The number and distribution of fungiform papillae and taste buds after lingual nerve injuries in cats.

Taste buds and the papillae that carry them are trophically dependent upon an intact innervation. The number and size of fungiform papillae and the number of taste buds on their surface were determined 3 months after crushing or sectioning the combined trunk of the chorda tympani and lingual nerves. After crushing there were fewer fungiform papillae but their size and number of taste buds were similar to that on the unoperated side. After nerve section there were considerably fewer recognizable fungiform papillae on the operated side and each carried a reduced number of taste buds.

Animals↗

Taste bud distribution in the rat pharynx and larynx.

The present study evaluated the number and distribution of taste buds in the rat pharynx and larynx using light microscopic techniques. The average number of taste buds in this region was 141.9 (S.D. = 29.4, n = 10). Pharyngeal and laryngeal taste buds could be grouped into three subpopulations: those associated with laryngeal structures (LA), the nasopharynx (NP), or the palatopharyngeal eminence (PE). Laryngeal taste buds were most numerous (mean = 103.2, S.D. = 23.6). They were observed on the laryngeal surface of the epiglottis and extended caudally along the aryepiglottic folds, reaching peak density at the caudal extreme of the folds. Further caudally, when the larynx and rostral esophagus separated, taste bud density decreased sharply. Fewer taste buds were associated with the NP (mean = 31.9, S.D. = 13.7) or PE (mean = 6.8, S.D. = 4.0) and taste bud density in these subpopulations peaked at the anterior border of the nasopharyngeal hiatus. Taste buds in the rat larynx and pharynx account for 10% all taste buds in this species, a proportion similar to that accounted for by taste buds on the soft palate. Taste buds in this region appear to be ideally situated for protecting the airway during accidental aspiration of food or fluids. Most pharyngeal and laryngeal taste buds are unlikely to be involved in triggering normally occurring swallows, although they could be stimulated as the bolus is propelled from the oral cavity to the esophagus.

Animals↗

Effects of zinc deficiency on the vallate papillae and taste buds in rats.

BACKGROUND AND PURPOSE: Zinc deficiency is associated with multiple clinical complications, including taste disturbance, anorexia, growth retardation, skin changes, and hypogonadism. We investigated the zinc-deficiency-induced morphologic changes in the vallate taste buds of weanling and young adult male Wistar rats. METHODS: A total of 24 weanling and 30 young adult rats were used. Each age group was further divided into a control group fed a zinc-adequate (50 ppm) diet, a zinc-deficient (< 1 ppm) diet group, and a zinc-adequate pair-fed group who were fed the same amount of food as that taken by the zinc-deficient group. Weanling rats were fed for 4 weeks and young adult rats were fed for 6 weeks. The morphometry and morphologic changes of vallate taste buds were analyzed using light and transmission electron microscopy. RESULTS: Light microscopy revealed no significant difference in papilla size and morphology among the various groups. In both weanling and young adult rats in the zinc-deficient diet and pair-fed groups, the number of taste buds per papilla (per animal) and the average profile area of the taste bud were significantly smaller than those of the corresponding controls (p < 0.05). Ultrastructural changes were seen only in the taste buds of weanling rats fed the zinc-deficient diet, with derangement of the architecture of the taste bud and widening of the intercellular space between taste bud cells. The proportion of type I taste bud cells in the taste buds of weanling rats fed the zinc-deficient diet decreased from 59% to 39%, and that of type II taste bud cells decreased from 25% to 12%. No obvious changes in the ultrastructure of type III taste bud cells were observed. CONCLUSIONS: The main effects of zinc deficiency in weanling and young adult rats and in adequate diet pair-fed rats were changes in the number and size of taste buds, and fine structure changes in the taste bud cells, especially during the accelerated growth stage after weaning.

Aging↗

Immunocytochemistry of gamma-aminobutyric acid, glutamate, serotonin, and histamine in Necturus taste buds.

Little information is currently available about which neurotransmitters are involved in signal processing in the peripheral sensory organs of taste, taste buds. Synaptic contacts between taste cells and sensory axons have long been known to exist, but what substances are active at these synapses is not known. Our objective in this study was to test for the presence of the neurotransmitter candidates, GABA, glutamate, serotonin, and histamine in taste buds of Necturus maculosus. Light microscopic immunocytochemical techniques were used to investigate the location of these substances in taste buds and surrounding epithelium. GABA and glutamate were detected in nerve fibers that innervate the taste buds, and, to a substantially lesser extent, in fine, varicose axons that penetrated the surrounding nontaste epithelium. Serotonin immunostaining was strong in basal cells in frog taste discs but was only faintly detected in Necturus taste buds. Histamine was not detected at all in taste buds. We conclude that amino acid neurotransmission may be involved in taste mechanisms and that monoamines may also play a role in chemosensory transduction in the taste bud. On the basis of our inability to detect histamine with immunocytochemical techniques, we conclude that this substance is unlikely to be a major neurotransmitter in Necturus taste buds.

Animals↗

Transcellular labeling by DiI demonstrates the glossopharyngeal innervation of taste buds in the lingual epithelium of the axolotl.

Innervation of the axolotl lingual epithelium by the glossopharyngeal nerve was examined to reveal its sensory target cells. The carbocyanine dye diI was applied to the nerve stump in the tongue fixed with paraformaldehyde. After a diffusion period of several months, the tongues were examined with a conventional epifluorescence microscope and a confocal laser scanning microscope (LSM) in wholemounts or preparations sectioned with a vibratome. Beneath the epithelium the labeled nerve fibers spread horizontally to form a meshwork of fibers, from which fascicles of fibers extended upward perpendicularly to the epithelium to innervate taste buds. Numerous taste buds were labeled by possible transcellular diffusion of diI. At the base of the taste bud, the nerve fibers branched and formed a basal plexus of fine fibers, on which numerous varicosities were seen. One or at most several taste cells were labeled in a taste bud. In the basal part of taste buds, the cell without an apical process, the basal cell, was also labeled. In the epithelium, between the taste buds, a few solitary cells were labeled. In some cases, a single fascicle of fibers innervating these cells was clearly shown by the LSM. In addition, fine fibers apparently formed free nerve endings in the epithelial cell layer. The results showed that the IX nerve innervated not only taste cells, but also presumed mechanosensory basal cells in the taste bud and the solitary cells of unknown function in the non-taste lingual epithelium. Afferent nerve responses to mechanical stimulation of the tongue may be explained by these non-taste cellular elements in the epithelium.

Ambystoma mexicanum↗

Amphibians provide new insights into taste-bud development.

Until recently, the predominant model of taste-bud development was one of neural induction: ingrowing sensory fibers were thought to induce taste-bud differentiation late in embryonic development. Recent experimental studies, however, show that the development of taste buds is independent of their innervation. In amphibian embryos, the ability to generate taste buds is an intrinsic feature of the oropharyngeal epithelium long before the region becomes innervated. These studies indicate that patterning of the oropharyngeal epithelium occurs during gastrulation, and suggest that taste buds or their progenitors play the dominant role in the development of their own innervation.

Amphibians↗

Keratin 19-like immunoreactivity in receptor cells of mammalian taste buds.

Three monoclonal antibodies, 4.62, LP2K and 170.2.14, were used to evaluate keratin 19-like immunoreactivity in gustatory epithelia. Keratin 19-like immunoreactivity was restricted to the intragemmal cells for all types of mammalian taste buds examined. These taste buds included fungiform, foliate and vallate taste buds in rat, gerbil and rabbit, and nasopalatine, epiglottal and palatine taste buds in rat. There was no keratin 19-like immunoreactivity in basal cells or in perigemmal cells lateral to the immunoreactive taste receptor cells. Denervation of the rat vallate papilla eliminated all taste buds, as well as all immunoreactive taste cells. That the immunoreactive material in the taste cells was keratin 19 was supported by the comparable staining of rat taste buds with each of three monoclonal antibodies specific for keratin 19. Furthermore, as predicted, these antibodies selectively stained luminal cells of rat bile ducts, bladder, salivary ducts, trachea, ureter and uterus. It was concluded that monoclonal antibodies against keratin 19 can usefully distinguish intragemmal taste receptor cells from keratinocytes, and from the perigemmal and basal cells of gustatory epithelia. Anti-keratin 19 antibodies may serve to identify differentiated taste cells in gustatory epithelia undergoing taste bud development, renewal, degeneration or regeneration.

Animals↗