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PubMed1992

Neural cell adhesion molecule of taste buds.

The distribution of the neural cell adhesion molecule (N-CAM) in the taste buds of adult and developing mice was studied by use of an immunocytochemical technique. In adult mice, nerve fibers and some elongated taste bud cells reacted with anti-N-CAM antibody. Immunoelectron microscopic observation showed that only type-III (gustatory) cells, which have afferent synaptic contacts with the nerves, were N-CAM positive in the taste buds. At day 0 after birth, taste buds of the developing mice first appeared in the circumvallate papillae, and N-CAM positive cells, which corresponded to type-III cells having afferent synapses, were found. This result suggests that N-CAM is involved in formation of the synaptic contacts between type-III cells and nerves in the taste buds.

Aging↗
PubMed1998

The role of innervation in the development of taste buds: insights from studies of amphibian embryos.

Amphibian embryos have long been model organisms for studies of development because of their hardiness and large size, as well as the ease with which they can be experimentally manipulated. These particular advantages have allowed us recently to test the role of innervation in the development of vertebrate taste buds using embryos of an aquatic salamander, the axolotl. The predominant model of taste bud genesis has been one of neural induction, in which ingrowing sensory neurites induce taste bud differentiation in the epithelium that lines the mouth and pharynx. However, when we prevented embryonic sensory neurons from contacting the oropharyngeal epithelium by using transplantation or tissue culture techniques, we found that taste bud differentiation was independent of nerve contact. Additionally, using similar types of experimental manipulations, we have recently shown that taste bud differentiation is not a result of interactions of the oropharyngeal epithelium with craniofacial mesenchyme. Surprisingly, we found that although taste bud genesis occurs very late in embryonic development, it is an intrinsic feature of the presumptive oropharyngeal epithelium extremely early, in fact as early as the completion of gastrulation. These data have prompted us to propose a new model for the development of amphibian taste buds: (i) The presumptive oropharyngeal epithelium is specified by the time gastrulation is complete; (ii) Subsequently, a distributed population of taste bud progenitors is set up within this epithelium via local cell-cell interactions. These progenitor cells give rise to taste buds, which are distributed throughout the mouth and pharynx. How widely applicable this model might be for the genesis of taste buds in other vertebrates remains to be seen. However, since it is likely that the taste system of axolotls more closely resembles the ancestral state from which both the amphibian and mammalian taste systems have evolved, it is possible that many of the same developmental mechanisms that give rise to amphibian taste buds are also used to generate the receptor organs in mammals.

Ambystoma↗
PubMed2003

The time course of taste bud regeneration after glossopharyngeal or greater superficial petrosal nerve transection in rats.

We previously have published data detailing the time course of taste bud regeneration in the anterior tongue following transection of the chorda tympani (CT) nerve in the rat. This study extends the prior work by determining the time course of taste bud regeneration in the vallate papilla, soft palate and nasoincisor ducts (NID) following transection of either the glossopharyngeal (GL) or greater superficial petrosal (GSP) nerve. Following GL transection in rats (n = 6 per time point), taste buds reappeared in the vallate papilla between 15 and 28 days after surgery, and returned to 80.3% of control levels (n = 12) of taste buds by 70 days postsurgery. The first appearance and the final percentage of the normal complement of regenerated vallate taste buds after GL transection resembled that seen previously in the anterior tongue after CT transection. However, in the latter case, regenerated taste buds reached asymptotic levels by 42 days after surgery, whereas within the time frame of the present study, a clear asymptotic return of vallate taste buds was not observed. In contrast to the posterior (and anterior) tongue, only 25% of the normal complement of palatal taste buds regenerated by 112 days and 224 days after GSP transection (n = 9). The difference in regenerative capacity might relate to the surgical approach used to transect the GSP. These experiments provide useful parametric data for investigators studying the functional consequences of gustatory nerve transection and regeneration.

Animals↗
PubMed1965

Renewal of cells within taste buds.

Colchicine blocks mitotic division of the epithelial cells surrounding the taste bud of the rat tongue. Response to chemical stimulation decreases 50 per cent 3 hours after colchicine injection as measured by recording the electrical activity from the taste nerve bundle. Radioautography, using tritiated thymidine, shows that those epithelial cells surrounding the taste bud divide and that some of the daughter cells enter the taste bud and slowly move toward the center. The life span of the average cell is about 250 +/- 50 hours, although some cells have a much shorter and others a much longer life span. These studies suggest that the cells within the taste bud, as well as the nerves, undergo considerable change with time. Corresponding changes in function are considered.

Animals↗
PubMed1997

Occurrence of subtypes of gustatory cells in cat circumvallate taste buds.

Transmission-electron microscopy of cat taste buds confirmed that, as in other mammals, each taste bud comprised four distinct types of cells: Type I, Type II, Type III (gustatory), and Type IV (basal) cells. Gustatory cells made synaptic contacts with nerves to which synaptic vesicles were gathered. The following are the main findings on the cat gustatory cells: 1) The synaptic vesicles of gustatory cells were essentially all dense-cored in type; small clear vesicles, which usually are intermingled in other mammals, could not be found. 2) The vesicles were accumulated not only in the synaptic area but also in the basal cytoplasm. This implies endocrine and paracrine functions. 3) On the basis of the fine structure of the vesicles, two subtypes of gustatory cells were discriminated. A large part of the cells contained vesicles measuring about 180 nm in diameter, while a small part had smaller ones of about 100 nm in diameter. This is the first demonstration of a dual population of gustatory cells in mammals, suggesting different messenger substances utilized.

Animals↗
PubMed1990

Electron microscopic demonstration of lectin binding sites in the taste buds of the European catfish Silurus glanis (Teleostei).

Taste buds in the European catfish Silurus glanis were examined with electron microscopic lectin histochemistry. For detection of carbohydrate residues in sensory cells and adjacent epithelial cells, gold-, ferritin- and biotin-labeled lectins were used. A post-embedding procedure carried out on tissue sections embedded in LR-White was applied to differentiate between the sensory cells: The lectins from Helix pomatia (HPA) and Triticum vulgare (WGA) bound to N-acetyl-galactosamine and to N-acetylglucosamine residues occurring especially in vesicles of dark sensory cells. This indicates a secretory function of these cells. Most light sensory cells--with some exceptions, probably immature cells--, are HPA-negative. The mucus of the receptor field and at the top of the adjacent epithelial cells was strongly HPA-positive. Pre-embedding studies were performed in order to obtain information about the reaction of the mucus with lectins under supravital conditions. The mucus of the taste bud receptor field exhibited intensive binding to WGA, but not to the other lectins tested. Most lectins bound predominantly to the surface mucus of the nonsensory epithelium and to the marginal cells close to the receptor field. The strong lectin binding to mucins and the relatively weak lectin binding to cell surface membranes in pre-embedding studies suggest that the mucus possibly serves as a barrier which is passed selectively only by a small amount of lectins or lectin-carbohydrate complexes. Lectin-carbohydrate interactions may play a role in recognition phenomena on the plasmalemmata of the taste bud sensory cells. Recognition processes directed to bacteria or viruses should be considered as well.

Animals↗
PubMed1981

Regeneration of taste buds after reinnervation of a denervated tongue papilla by a normally nongustatory nerve.

Taste buds degenerate and disappear after transection of their sensory nerve supply, and they differentiate anew from epithelial cells (e.g., lingual) following regeneration of sensory but not motor or autonomic axons. A controversy exists as to whether only gustatory sensory nerves can cause buds to reform or whether any sensory nerve can perform this function. This issue arose because the results of cross-innervation studies revealed a specificity whereas grafting data demonstrated a nonspecificity. A retest of specificity in the cross-reinnervation situation was performed by reinnervating the denervated vallate papilla of adult rat tongue with a sensory branch of the vagus nerve that is not normally gustatory. It was found that taste buds disappeared and remained lost from acutely and chronically denervated papilla. However, some buds were found 90-100 days after reinnervation by the normally nongustatory vagus nerve branch. Transection of the regenerated vagus nerve resulted in the loss of innervation and the degeneration of taste buds from reinnervated papilla indicating that this nerve had supported buds. These results show that a normally nongustatory nerve can induce the formation of taste buds after its axons grow into appropriate tissue. It appears that the ability to support taste buds is a nonspecific, rather than a specific, property of sensory nerve.

Animals↗
PubMed1976

Biochemical studies of taste sensation. III. Preparation of a suspension of bovine taste bud cells and their labeling with a fluorescent probe.

A method to prepare suspensions of taste bud cells is described. Bovine circumvallate papillae, which contain most of the taste buds in this animal, are incubated in collagenase-containing medium and the epidermal sidewall tissue is then dissected from the inner gelatinous dermis. The sidewall tissue, which contains the taste buds, is gently homogenized by manual operation of an all-glass homogenizer with a loose-fitting pestle. The suspended material is separated on a discontinous Ficoll gradient (2%, 8%, 10%, 12% w/w). The material banding at the 8-2% interface is greatly enriched in spindle-shaped cells that are morphologically similar to taste bud cells as they appear in situ. These cells are not seen when the procedure is done with tissues devoid of taste buds, namely the upper surface of the circumvallate papilla or epithelium from the intermolar eminence. Fluorescence analysis indicates that the hydrophobic probe, 8-anilino-1-naphthalenesulfonate (ANS), binds to relatively nonpolar sites in the suspension. It is postulated that the probe is adsorbing onto the surface membrane of the cell. These preparations may be useful in studying specificity and transduction in taste sensation.

Anilino Naphthalenesulfonates↗
PubMed1981

Biogenic monoamines in developing taste buds of mouse circumvallate papillae.

After administration of monoamine precursors, developing taste buds of newborn and young mice were observed by means of electron microscopy and fluorescence histochemistry. Gustatory (type III) cells occurred in the primitive taste buds during stage 1 (0-1 day after birth). These cells had an immature type of afferent synaptic contacts with nerve terminals; however, no specific fluorescence was found in the taste buds after administration of 5-HTP or L-DOPA. During stage 2 (2-7 days), mature types of afferent synapses, taste pores, type I cells and type II cells appeared in the taste buds, and fluorescent cells also appeared following treatment of 5-HTP or L-DOPA. During stage 3 (14-21 days), the gustatory cells underwent ultrastructural changes following injection of 5-HTP; i.e. small dense-cored vesicles (30-60 nm) appeared scattered throughout the cytoplasm and were found to intermingle with small clear vesicles accumulated at the presynaptic membranes of afferent synapses, and the electron densities of large dense-cored vesicles (80-100 nm) were elevated as compared with those of untreated mice. Consequently the ability of gustatory cells to take up amine-precursors started simultaneously with the formation of taste pores and mature afferent synapses between the gustatory cells and the sensory nerves.

5-Hydroxytryptophan↗
PubMed1973

Disc electrophoresis of extracts from the taste buds located in circumvallate papillae of rat tongues.

The epithelium of the circumvallate papillae of rat tongues was stripped off by treatment with 0.005% elastase in a state when many taste buds were present. The taste buds were isolated from the stripped epithelium by further treatment with 0.005% elastase and 0.08% trypsin. A protein which was thought to be characteristic of taste buds was found from semimicro disc polyacrylamide gel electrophoretic studies of the stripped epithelia with and without taste buds. This result was supported by micro disc polyacrylamide gel electrophoretic studies of isolated taste buds.

Animals↗
PubMed2003

Distribution of keratin 8-containing cell clusters in mouse embryonic tongue: evidence for a prepattern for taste bud development.

The initiation of the morphogenesis of gustatory papillae is independent of innervation. To address the question of whether taste bud formation is associated with gustatory papilla morphogenesis, we examined developing tongues in mouse embryos from embryonic day 11 to birth. Despite the smooth morphological appearance of the lingual dorsal surface at 13 days of gestation, we observed embryonic taste bud primordia as discrete collections of cytokeratin 8-positive and elongated cells in epithelial placodes in the anterior tongue. In subsequent stages until birth, cytokeratin 8 continues to be expressed in embryonic taste buds distributed in punctuate patterns at regular intervals along rows that are symmetrically located on both sides of the median sulcus in the dorsal anterior developing tongue. Embryonic taste buds were observed in the developing circumvallate papillae from 15.5 days of gestation until birth. The dorsal epithelium of the anterior tongue is not innervated when embryonic taste buds first occur. The increased numbers of embryonic taste buds in developing fungiform papillae until birth are not correlated with the neural invasion of the epithelium. Thus, taste buds occur prenatally more likely independently of the innervation.

Animals↗
PubMed1990

Keratin filaments of epithelial and taste-bud cells in the circumvallate papillae of adult and developing mice.

Keratin filaments of epithelial- and taste-bud cells in the circumvallate papillae of adult and developing mice were studied by immunocytochemistry using monoclonal antikeratin antibodies (PKK2 and PKK3) and by conventional electron microscopy. Elongated cells (type-I, -II, and -III cells) of the taste buds were stained by PKK3 antibody, which reacts with 45-kdalton keratin, whereas basal cells of the taste buds and surrounding epithelial cells showed negative staining with PKK3. Such PKK3-reactive cells occurred at 0 day after birth, when taste-buds first appeared in the dorsal surface epithelium of the papillae. Thus 45-kdalton keratin seems to be an excellent immunocytochemical marker for identifying taste-bud cells. Epithelial cells in all layers of the trench wall and basal layer cells of the dorsal surface contained densely aggregated bundles of keratin filaments that reacted with PKK2 antibody, but not with PKK3. In contrast, taste-bud cells and spinous and granular layer cells of the dorsal surface possessed loose aggregated bundles of filaments that reacted with PKK3, but not with PKK2. These results suggest that the aggregation and distribution pattern of keratin filaments may reflect differences in the keratin subtypes that comprise these filaments.

Actin Cytoskeleton↗
PubMed2007

Expression of T1Rs and gustducin in palatal taste buds of mice.

The palatal region of the oral cavity in rodents houses 100-300 taste buds and is particularly sensitive to sweet and umami compounds; yet, few studies have examined the expression patterns of transduction-related molecules in this taste field. We investigated the interrelationships between members of the T1R family and between each T1R and gustducin in palatal taste buds. Similar to lingual taste buds, T1R1 and T1R2 are generally expressed in separate palatal taste cells. In contrast to lingual taste buds, however, T1R2 and T1R3-positive palatal taste cells almost always coexpress gustducin, suggesting that sweet taste transduction in the palate is almost entirely dependent on gustducin. T1R1-positive palate taste cells coexpress gustducin about half the time, suggesting that other G proteins may contribute to the transduction of umami stimuli in this taste field.

Animals↗
PubMed1978

Histochemical observations on the taste buds of the marmosets (Callithrix jacchus and Callithrix penicillata).

The epithelial cells in the taste buds of C. jacchus and C. penicillata show a moderate amount of ribonucleic acid an a concentration of a PAS-positive diastase-resistant material at their apical part. These cells are devoid of UDPG-GT, phosphorylases, G-6-PA, alanyl aminopeptidase, leucine aminopeptidase, cholinesterase and MAO; they present a weak reaction of F-1, 6-P Ald, LDH, SDH, MDH, cytochrome oxidase, beta-OHBDH, nonspecific esterase and acid phosphatase and a stronger reaction to ADH, NADPH2-TR, ATPases, alpha-GPDH, alkaline phosphatase, 5-nucleotidase and GDH. Although some enzymes (alkaline phosphatase, 5-nucleotidase and ATPases) have an almost uniform reactivity by the several taste buds, the other ones react with a lesser intensity in the smaller uniform reactivity by the several taste buds, the other ones react with a lesser intensity in the smaller taste buds of the fungiform papillae. As a rule the apical part of the cells shows a stronger enzymatic reactivity. The taste buds of the marmosets are penetrated by acetylcholinesterase positive nerve fibers whereas the autonomic ganglia in the connective tissue contain both-acetyl and butyrylcholinesterase.

Adenosine Triphosphatases↗
PubMed2005

Expression of glial cell line-derived neurotrophic factor (GDNF) and GDNF family receptor alpha1 in mouse taste bud cells after denervation.

Glial cell line-derived neurotrophic' factor (GDNF) has been isolated as a neurotrophic factor that affects the survival and maintenance of central and peripheral neurons. Using immunocytochemical methods, we examined whether the taste bud cells in mouse circumvallate papillae after transection of the glossopharyngeal nerves expressed GDNF and its receptor, GDNF family receptor alpha1 (GFRalpha1). By 5 and 10 days after denervation, the number of taste buds had decreased markedly; however, the remaining taste bud cells still expressed GDNF and GFRalpha1. By 14 days after denervation, most of the taste buds had disappeared and GDNF- and GFRalpha1-immunoreactive cells were not seen. By 4 weeks after denervation, numerous TrkB-immunoreactive nerve fibers had invaded the papilla and a few taste buds expressing GDNF and GFRalpha1 had regenerated. Thus, GDNF- and GFRalpha1-immunoreactive taste bud cells after denervation vanished following the disappearance of the taste buds and reappeared at the same time as the taste buds reappeared.

Animals↗
PubMed1975

On the ultrastructure and permeability of taste buds of the marine teleost Ciliata mustela.

The abundant taste buds of the barbels and free fin rays of the five bearded rockling, Ciliata mustela contain an average of 100-150 cells, falling into two types. Tubule-containing cells ('t-cells'), tentatively identified as receptor cells, and each surrounded by fibril-containing cells ('f-cells') in the central part of the bud. t-Cells also occur in two concentric shells separated by indifferent epithelial cells at the periphery of the bud. f-Cells are characterized by their concentrations of fine fibrils, and by granules or vesicles of 180-190 mmu diameter. The 100 or so receptor cells in a taste bud are innervated by some 250 axons. Lanthanum penetrates more deeply into the extracellular space of taste buds than into the extracellular space of the general epithelium, perhaps indicating that a greater area than the mere protruding tip of receptor cells may be accessible to chemical stimulation. Degenerating cells may provide an important route of entry for such external agents.

Animals↗
PubMed1980

Distribution of taste buds on the lips and inside the mouth in a minnow, Pseudorasbora parva.

The distribution and abundance of taste buds were quantitatively examined by observing silver impregnated serial sections. The taste buds were widely dispersed on the skin, the lips, the mucosa in the oro-pharyngeal cavity, the esophagus, and the branchial apparatus. The great majority of them was found on the lips and inside the mouth. The external buds were concentrated especially on the outer lips and the adjacent skin, while their number diminished in a caudal direction. Very few were found on the scaled skin. The total number of external buds in a specimen of 6 cm in length was 1,486. The number of taste buds inside the mouth was 6,600. On the inner lips and the palatal organ densities were found to reach over 140 per mm2. High concentrations of taste buds were also found on the gill arches and rakers. These taste buds varied to some extent in size and shape, depending on the thickness of the epithelial layer. It is suggested that the minnow may use the lips, gills and palatal organ as its main taste organs.

Animals↗
PubMed1999

Expression of Mash1 in basal cells of rat circumvallate taste buds is dependent upon gustatory innervation.

Mash1, a mammalian homologue of the Drosophila achaete-scute proneural gene complex, plays an essential role in differentiation of subsets of peripheral neurons. In this study, using RT-PCR and in situ RT-PCR, we investigated if Mash1 gene expression occurs in rat taste buds. Further, we examined dynamics of Mash1 expression in the process of degeneration and regeneration in denervated rat taste buds. In rat tongue epithelium, Mash1 gene expression is confined to circumvallate, foliate, and fungiform papilla epithelia that include taste buds. In taste buds, Mash1-expressing cells are round cells in the basal compartment. In contrast, the mature taste bud cells do not express the Mash1 gene. Denervation and regeneration experiments show that the expression of Mash1 requires gustatory innervation. We conclude that Mash1 is expressed in cells of the taste bud lineage, and that the expression of Mash1 in rat taste buds is dependent upon gustatory innervation.

Animals↗