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Fine structure of the canine taste bud with special reference to gustatory cell functions.

Canine taste buds were observed by transmission electron microscopy with special reference to the gustatory cell function. The cells forming the taste bud were divided into five types: four of these corresponded to the previously reported Types I, II, III and IV (basal cell); another type, a slender, immature-looking cell located at the outermost layer, was identified as the peripheral cell known in the rat. The Type I cell was supportive in nature, located between other cell types and enveloping nerve fibers. This cell apically secretes dense mucous substances. The Type II cell was in broad contact with nerve fibers and constantly contained a subsurface cistern beneath them. In the dog, this cell was characterized by a large supranuclear Golgi apparatus. The Type III cell was, as in other animals, regarded as the gustatory cell since it made synaptic contacts with nerves and contained synaptic vesicles. Numerous large corded vesicles were intermingled with some small clear vesicles. In addition to accumulating to the synaptic areas, the vesicles often filled the base of the cell, which was irregularly thickened, showing a more or less extensive contact with the basement membrane. In this area, numerous large cored vesicles approached the cell base. Blood capillaries close to the base of the taste bud were fenestrated in the endothelium. These findings supported a hypothesis that the transmitters may be released from the base of the gustatory cell and possibly exert paracrine and endocrine (hemocrine) effects. The salivary gland of Ebner was suggested as one such target. The nerve terminals on the Type III cell also contained synaptic vesicles, suggesting a reciprocal nature of the synapses. The Type IV cell was the immature basal cell which has been established in other species; mitosis could be seen in this cell.

Animals↗

Isolation of taste buds from the foliate papillae of the rabbit.

A method to isolate taste buds from the foliate papillae of the rabbit tongue is described. The method comprises (a) separation of the epidermis from the dermal layer after treatment with dilute acetic acid, and (b) mechanical removal of the taste buds from the epithelium with the use of a surgical needle. The procedure yields taste buds that are morphologically well preserved, and in quantities sufficient to enable a detailed biochemical characterization. Preliminary tests have shown the taste buds to have biochemical properties clearly distinct from those of the adjacent epithelium. The method may provide a basis for studying the molecular mechanism of taste perception in greater detail.

Acetates↗

Innervation of taste buds in the canine larynx as revealed by immunohistochemistry for the various neurochemical markers.

The distribution and innervation of the canine laryngeal taste buds were observed using immunohistochemistry with antibodies against protein gene product 9.5 (PGP 9.5) and neurofilament protein (NFP). We also observed the immunohistochemical distribution of serotonin, tyrosine hydroxylase (TH) and various neuropeptides including calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal peptide (VIP), galanin, methionine enkephalin (ENK) and neuropeptide Y (NPY). The taste buds in the canine larynx were densely distributed in the mucosa at the basal portion of the epiglottis and cuneiform process of the arytenoid cartilage. The taste cells were immunoreactive for PGP 9.5 and serotonin. The nerve fibers with immunoreactivity for PGP 9.5 in the taste buds were observed in the perigemmal region and intra- and subgemmal plexuses, and these were classified into two types based on their diameter. The thick nerve fibers corresponded to the fibers immunoreactive for NFP, while the thin nerve fibers corresponded to the fibers immunoreactive for TH and various neuropeptides. Numerous nerve fibers immunoreactive for SP and CGRP were observed in the perigemmal region, and intra- and subgemmal plexuses. A few galanin- and ENK-immunoreactive nerve fibers were also observed in the taste buds, whereas NPY-immunoreactive nerve fibers were noted beneath them. All peptide-containing fibers except for VIP-immunoreactive nerves were situated in the subgemmal regions. In conclusion, the multiple innervation to the laryngeal taste buds were documented. Thick nerve fibers are likely to be irritant receptors, while thin varicose nerve fibers seem to regulate taste buds themselves. The laryngeal taste buds may be among the important structures which are sensitive to exogeneous chemical and/or mechanical stimuli, for the protection of the airway and the regulation of the respiratory function.

Animals↗

Structure of bovine fungiform taste buds and their immunoreactivity for gustducin.

The taste buds of bovine fungiform papillae were studied by light and electron microscopy using both histological and immunohistochemical methods. The taste buds existed in the epithelium of the apical region of the papillae. By electron microscopy, two types of taste cells, namely type I and type II cells, could be classified according to the presence of dense-cored vesicles, the cytoplasmic density and the cell shape. Type I cells were thin, had an electron-dense cytoplasm containing dense-cored vesicles, and possessed long thick apical processes in the taste pore. Type II cells were thick, had an electron-lucent cytoplasm containing many electron-lucent vesicles, rather than dense-cored vesicles, and possessed microvilli in the taste pore. Immunohistochemical staining with an antiserum against gustducin was investigated by both light and electron microscopy using the avidin-biotin complex (ABC) method. Some, but not all, of the type II cells exhibited gustducin immunoreactivity, whereas none of the type I cells showed any immunoreactivity.

Animals↗

Taste bud adenosine -3'5'-monophosphate phosphodiesterase: activity, subcellular distribution and kinetic parameters.

Higher activity of cyclic adenosine 3',5'-monophosphate (cAMP) phosphodiesterase (PDE) was found in homogenates from bovine circumvellate papillae bearing taste buds compared to activity in homogenates from areas surrounding these papillae in which no taste buds were present. With progressive purification of these homogenates cAMP PDE activity increased in the taste bud enriched fractions relative to that measured in the non-taste bud bearing epithelial tissue. The highest levels of cAMP PDE activity were measured in those taste bud fractions in which purification was greatest. Kinetic studies in both taste bud derived and control tissues showed two apparent Km values, one relatively high, the other, lower. cAMP PDE activity of taste bud membranes was enhanced by Mg++, Mn++ and imidazole and inhibited by ethylene-bis (beta-aminoethylether) N,N-tetra-acetic acid (EGTA), isobutyl methyl xanthine (IBMX), theophylline, and cyclic guanosine 3',5'-monophosphate (cGMP). The possible role for cAMP PDE in taste function is discussed.

3',5'-Cyclic-AMP Phosphodiesterases↗

Human blood group antigen H is not the specific marker for type I cells in the taste buds.

We examined the localization of human blood antigen H (AbH) and its correlation with other cell type markers in the taste buds of circumvallate papillae of the adult rat. Immunoreactivity for AbH was localized in the membrane of two cell populations in the taste buds: in spindle-shaped cells extending from base to the apical portion of the taste buds as well as in round-shaped cells at the basal portion of the taste buds. Quantitative analysis revealed that approximately 47.8%, 24.4%, and 14.6% of cells within the taste buds displayed AbH-, alpha-gustducin- or protein gene product 9.5 (PGP 9.5)-immunoreactivity, respectively. Approximately 16.3% and 6.6% of AbH-immunoreactive taste bud cells displayed alpha-gustducin- or PGP 9.5-immunoreactivity, respectively. Although previous studies proposed that AbH immunoreactivity was specific for type I cells (dark cells or supporting cells), the present results indicate that AbH immunoreactivity is also present in some type II cells (alpha-gustducin immunoreactive cells) and type III cells (PGP 9.5-immunoreactive cells).

ABO Blood-Group System↗

Differential expression of brain-derived neurotrophic factor and neurotrophin 3 mRNA in lingual papillae and taste buds indicates roles in gustatory and somatosensory innervation.

Although many studies have demonstrated the dependency of taste bud function and/or survival on intact innervation, relatively few have dealt with the development of taste bud innervation. Using in situ hybridization histochemistry, we show that brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT3) mRNA are expressed in a specific pattern in the taste buds, tongue papillae, and lingual epithelium during development and that expression persists into adulthood. BDNF mRNA is expressed in a fraction of the taste cells of the developing and adult taste buds in rats, showing different labeling intensities among the labeled cells. NT3 and mRNA seems to be located in areas other than those where BDNF mRNA is expressed, mainly in the superior epithelial surfaces of circumvallate papillae, the outer surface epithelium of foliate papilae, the superior surface and the lateral epithelium of the fungiform papillae, and the epithelium of the filiform papillae. NT3 mRNA labeling is also observed among muscle and connective tissue of the tongue. The morphological appearance, expression of NT3 mRNA, and ramification of nerve fibers in defined epithelial structures in the posterior wall of the anterior filiform papillae suggest the existence of a mechanosensory apparatus in these papillae. Nerve growth factor and neurotrophin 4 probes did not give rise to selective labeling in tongue, although their presence cannot be totally excluded. Based on present and prior studies, we suggest that BDNF is needed during initiation and for maintenance of gustatory innervation of taste buds and gustatory papillae and that NT3 is mainly needed for somatosensory innervation of the tongue.

Animals↗

Taste bud expression of human blood group antigens.

Some human blood group antigens are expressed by rodent epithelial cells at different stages of differentiation. Since adult taste cells are continually replaced throughout life, we investigated the expression of the H, B, A and Lewisb blood group determinants by cells of the rat fungiform, foliate and vallate papillae. We employed antibodies against the trisaccharide structures of the H, B, and A blood group antigens and against the Lewisb blood group epitope in studies of normal and denervated taste buds. The antibody against the H antigen reacted with the majority of cells in all taste buds and with cells in the spinous layer of the tongue epithelium. The B antigen was expressed by the majority of taste cells but not by other epithelial cells. The expression of the A antigen was significantly less in the fungiform taste buds than in the vallate or foliate taste buds. The A antigen was also abundantly expressed in the acini of the lingual salivary glands. The Lewisb epitope was expressed by a subset of cells in taste buds of the fungiform, foliate and vallate papillae. Taste buds are trophically dependent upon gustatory nerve innervation. Transection of the chorda tympani or the IXth nerve resulted in the loss of expression of these molecules from the gustatory epithelium, indicating that they are expressed only on differentiated taste cells. The blood group antigens are lactoseries carbohydrates; they are differentially expressed in developing cochlear hair cells and olfactory neurons and may play roles in cell-cell recognition, adhesion, and other interactions important in the developing nervous system. They could have similar functions in the taste and olfactory systems, where the receptors are continually renewed and new synapses between the receptors and their neural targets continually form.

ABO Blood-Group System↗

The effect of beta-bungarotoxin, or geniculate ganglion lesion on taste bud development in the chick embryo.

Chick taste bud (gemmal) primordia normally appear on embryonic day (E) 16 and incipient immature, spherical-shaped buds at E17. In ovo injection of beta-bungarotoxin at E12 resulted in a complete absence of taste buds in lower beak and palatal epithelium at developmental ages E17 and E21. However, putative gemmal primordia (solitary clear cells; small, cell groupings) remained, lying adjacent to salivary gland duct openings as seen in normal chick gemmal development. Oral epithelium was immunonegative to neural cell adhesion molecule (NCAM) suggesting gemmal primordia are nerve-independent. Some NCAM immunoreactivity was evident in autonomic ganglion-like cells and nerve fibers in connective tissue. After unilateral geniculate ganglion/otocyst excision on E2.5, at developmental ages E18 and posthatching day 1, approximately 12% of surviving ipsilateral geniculate ganglion cells sustained approximately 54% of the unoperated gemmal counts. After E18, proportional stages of differentiation in surviving developing buds probably reflect their degree of innervation, as well as rate of differentiation. Irrespective of the degree of geniculate ganglion damage, the proportion of surviving buds can be sustained at the same differentiated bud stage as on the unoperated side, or may differentiate to a later bud stage, consistent with the thesis that bud maturation, maintenance, and survival are nerve-dependent.

Animals↗

Taste buds and neuronal markers in patients with chronic renal failure.

OBJECTIVE: To study the number of taste buds and, with the use of specific markers for peripheral nervous tissue, to study the neuronal pattern in taste buds from 36 patients with chronic renal failure (CRF), 19 renal transplant recipients, and 40 healthy subjects. Of the patients with CRF, 17 patients had not started dialysis, 12 patients were on peritoneal dialysis, and 7 patients were on hemodialysis. DESIGN: From all subjects, two or three fungiform papillae were collected from the anterior part of the tongue. Cryostat sections were cut and inspected under light microscopy to determine the presence of taste buds. The sections were subsequently incubated with primary rabbit antibodies against protein gene product 9.5, substance P, and nerve growth factor receptor. RESULTS: Using these antibodies, no differences between the groups were observed. However, patients with CRF had fewer taste buds than control subjects. CONCLUSION: No immunohistochemical differences were observed between patients with CRF and healthy controls. However, patients with CRF had significantly fewer fungiform taste buds, suggesting an important factor contributing to the well-known impairment of taste acuity in this patient group.

Adult↗

Neurochemical markers of human fungiform papillae and taste buds.

The presence of distribution of several neurochemical markers in human fungiform papillae and taste buds were investigated by the immunohistochemical technique. The gustatory cells of the taste buds are in synaptic contact with sensory nerve endings, and considering the taste buds strictly as specialized sensory organs, the amounts and distribution of some of the neurochemical markers were different to what we expected. For example, few structures showed immunoreactivity to the tachykinins substance P (SP), calcitonin gene-related peptide (CGRP), and neurokinin A (NKA) also for the peptides vasoactive intestinal polypeptide (VIP), neuropeptide tyrosine (NPY) and galanin, low amounts of immunoreactivity occurred. On the other hand, using antibodies to protein gene product 9.5 (PGP 9.5), protein S-100, and glutamate, numerous nerve fibres and/or immunoreactive cells were found in the fungiform papillae, in the epithelium, in the connective tissue and around blood vessels, as well as in or near taste buds. Incubation with the antibodies against somatostatin, enkephalin, bombesin, peptide histidine isoleucine amide (PHI), cholecystokinin (CCK)/gastrin and dopamine-beta-hydroxylase (DBH) was negative for the fungiform papillae. In conclusion, the present study has shown several immunoreactive structures using antibodies against certain neurochemical markers. Further investigations will hopefully correlate these morphological findings with functional taste perception data. Future studies of patients with taste disorders or other pathological changes correlated with taste and tongue will also be of utmost importance.

Antibodies↗

Neurotrophin receptors in taste buds of adult zebrafish (Danio rerio).

TrkB plays crucial roles in the development and maintenance of taste buds in mammals. In this study we investigated the presence and cell localization of Trks (TrkA, TrkB and TrkC) in taste buds of the zebrafish (Danio rerio) using Western blot and immunohistochemistry. Proteins of 140 and 145 kDa, identified as full-length TrkA and TrkB, were detected. Conversely, the anti-TrkC antibody recognized a protein lower than expected (100 kDa). In agreement with these results the sensory cells of taste buds, displayed TrkA- and TrkB-like, but not TrkC-like, immunoreactivity. TrkA and TrkB co-existed in the same taste buds, but remains to be clarified whether or not they are co-expressed in the same cells. Present results demonstrate that as for mammals neurotrophins might play a role in sensory cells of the teleostean taste buds.

Animals↗

Morphological changes in taste buds of the fungiform papillae after a single dose of x-ray irradiation in rats.

The qualitative and quantitative changes occurring in the taste buds of the fungiform papillae of male Sprague-Dawley rats (aged 51-125 days old) after x-ray irradiation were studied. A single dose of 2000 Roentgen (R) was delivered and observations were made at 0, 3, 7, 14, 21 and 30 days after x-ray irradiation. The changes in taste bud morphology were interpreted first as degenerative and then as regenerative at various stages of the study. The degenerative process appeared at about the third day and reached a peak at 7 days. Regeneration then began after a further week and the taste buds appeared mature at about 30 days. While the number of fungiform papillae remained constant throughout the study, there was a marked loss of taste buds on these papillae at 7 days. At 14 days, the number of taste buds increased from this low level, and by 30 days the number approached the control values. Initial taste bud loss could be attributed to a direct irradiation damage leading to taste bud cell desquamation combined with a concomitant mitotic inhibition of the stratum germinativum.

Animals↗

Temporal changes in NCAM immunoreactivity during taste cell differentiation and cell lineage relationships in taste buds.

Neural cell adhesion molecule (NCAM) is a type III cell marker in the taste buds. In order to clarify the cell type of Mash1-expressing cells in taste buds, expression of NCAM was examined in Mash1-expressing taste cells of adult mice in comparison with gustducin- and T1r3-expressing cells, using a combination of NCAM immunohistochemistry and in situ hybridization. About 98% of Mash1-expressing cells were NCAM immunopositive (IP), suggesting that Mash1-expressing cells should be categorized as type III cells. Unexpectedly, small subsets of gustducin- and T1r3-expressing cells were also found to be NCAM-IP, contradicting previous immunohistochemical studies in rats, in which gustducin-IP cells were observed specifically in type II cells, which do not have NCAM immunoreactivity. Examinations of developing taste buds showed temporal changes in the ratio of NCAM-IP cells in gustducin- and T1r3-expressing cells; the ratio of NCAM-IP cells in these gene-expressing cells were approximately 90% at 0.5 days after birth and decreased markedly during development. In contrast, the majority of Mash1-expressing cells showed constant NCAM immunoreactivity throughout development. In addition, BrdU-labeling experiments showed that the differentiation of Mash1-expressing cells precedes those of gustducin- and T1r3-expressing cells in taste buds of adult mice. These results suggest that T1r3- and gustducin-expressing cells are NCAM-IP at the beginning of cell differentiation, and that NCAM immunoreactivity in gustducin- and T1r3-expressing cells might remain from the previous developmental stage expressing Mash1.

Animals↗

[Changes of taste bud and fungiform papillae after 60Co radiation in rat].

OBJECTIVE: To observe the morphological changes and the regenerating ability of the fungiform papillae and taste buds after 60Co radiation with clinical doses in rats. METHODS: The heads, faces and necks of 30 SD rats were radiated with a large dose and one time of 60Co in the clinical radiation. The general living condition and the number and shape of the fungiform papillae and taste buds of the tongues were observed after the radiation in rats. RESULTS: In the group of 60Co radiation, the animals had wilting, decreasing appetite, losing weight. The heads, faces and necks of animals appeared redness, peeling of hair, increasing of secretions in 5 days after the 60Co radiation. The changes reached the summit in 10 days and the general living condition of the animals recovered in 60 days. The fungiform papillae and taste buds of the animals appeared degeneration, atrophy and collapsing in 5 days after the 60Co radiation. The injuries reached the summit in 10-20 days and the fungiform papillae and taste buds regenerated partially, and the some atrophied fungiform papillae and taste buds were not regenerated in 60 days. CONCLUSION: The damage to fungiform papillae and taste buds of tongue following the 60Co radiation with the clinical doses was very serious. The damaged fungiform papillae and taste buds can regenerate partially, but not completely.

Animals↗

Sugar binding to purified fractions from bovine taste buds and epithelial tissue. Relationships to bioactivity.

Binding of various sugars was compared in purified subfractions of taste buds isolated from bovine circumvallate papillae and of non-taste bud-bearing epithelium isolated from tissue surrounding these papillae. Binding of 14C-labeled sugars was greater in purified subfractions obtained from taste bud than from non-taste bud-bearing tissue and was, in general, greater in those taste bud subfractions in which a greater membrane purification was achieved. Binding specificity of the 14C-labeled sugars sucrose, fructose, glucose and of 14C-labeled cyclamate and saccharine was measured by competition of each 14C-labeled sugar or synthetic sweetener with its unlabeled homologous sugar in P4(B) taste bud subfractions; this binding, as shown for sucrose, was reversible and temperature dependent. Essentially no competition of the 14C-lageled sugars sucrose, fructose, glucose or 14C-labeled cyclamate and saccharine by their respective unlabeled homologues occurred in epithelial tissue P4(B) subfractions; this binding was not reversible. Binding specificity was further observed by the competition of 14C-labeled sucrose, fructose and glucose with each unlabeled sugar for binding sites on P4(B) taste bud subfractions; unlabeled sucrose was more effective in competing with each 14C-labeled surgar than was unlabeled fructose or glucose. The relatively non-sweet sugar lactose did not compete with 14C-labeled lactose in P4(B) subfractions from either taste bud or non-taste bud-bearing epithelial tissue. Binding of 14C-labeled sucrose in purified P4(B) bud subfractions was inhibited by increased concentrations of unlabeled sucrose, phospholipase C, neuraminidase, EDTA, NaCl and urea. Dissociation constants for sugar or synthetic sweetener binding were low (approx. 10(-3) M) but in a rank order (sucrose greater than fructose greater than glucose greater than saccharine) consistent with preference and electrophysiological responses in cow. The cow is behaviorally indifferent to saccharine and lactose consistent with the data obtained in the present study.

Animals↗

Temporal and spatial patterns of tenascin and laminin immunoreactivity suggest roles for extracellular matrix in development of gustatory papillae and taste buds.

Gustatory papillae are complex organs that are composed of 1) an epithelium, 2) specialized sensory cells within the epithelium (the taste buds), 3) a broad connective core, and 4) sensory innervation. During papilla development, cells in the various tissue compartments must divide, aggregate, detach, migrate, and reaggregate in relation to each other, but factors that regulate such steps are poorly understood and have not been extensively studied. All of these processes potentially require participation of the extracellular matrix. Therefore, we have studied temporal and spatial patterns of immunoreactivity for two extracellular matrix molecules, tenascin and laminin, in the developing fungiform and circumvallate papillae of fetal, perinatal, and adult sheep tongue. To determine relations of tenascin and laminin to sensory innervation, we used an antibody to growth-associated protein (GAP-43) to label growing nerves. Immunocytochemical distributions of tenascin and laminin alter during development in a manner that reflects morphogenesis rather than histologic boundaries of the taste papillae. In early fungiform papillae, tenascin immunoreactivity is very weak within the mesenchyme of the papilla core. However, there is a subsequent shift to an intense, restricted localization in the apical papilla core only--directly under taste bud-bearing regions of the papilla epithelium. In early circumvallate papillae, tenascin immunoreactivity is patchy within the papilla core and within the flanking, nongustatory papillae. Later, immunoreactivity is restricted to the perimeter of the central papilla core, under epithelium that contains developing taste buds. In fungiform and circumvallate papillae, the shift in tenascin immunolocalization is associated with periods of taste bud formation and multiplication within the papilla epithelium and with extensive branching of the sensory innervation in the papilla apex. Laminin immunoreactivity, although it is continuous throughout the basement membrane of general lingual epithelium, is interrupted in the epithelial basement membrane of early fungiform and circumvallate papillae in regions where taste buds are forming. The breaks are large in young fetuses, when taste buds first develop, and are evidenced later as punctate disruptions. Heparan sulfate proteoglycan immunoreactivity confirms that these are basement membrane discontinuities. GAP-43 label coincides with innervation of the papilla core and is most extensive in regions where tenascin immunoreactivity is weak or absent. GAP-43 immunoreactivity is also found in early taste buds: Later, it is extensive within more mature multiple taste buds, presumably in relation to synaptogenesis. We propose that tenascin has a role in promoting deadhesion of cells in the papilla epithelium during periods of taste bud formation and multiplication. Discontinuities in the epithelial basement membrane under developing taste buds, indicated with laminin and heparan sulfate proteoglycan immunoreactivity, may interact to facilitate taste bud morphogenesis and multiplication, to permit access of papilla innervation to the forming taste buds, and/or to allow epithelial/mesenchymal interactions during papilla and taste bud development.

Animals↗

Mouse taste buds use serotonin as a neurotransmitter.

Synapses between gustatory receptor cells and primary sensory afferent fibers transmit the output signal from taste buds to the CNS. Several transmitter candidates have been proposed for these synapses, including serotonin (5-HT), glutamate, acetylcholine, ATP, peptides, and others, but, to date, none has been unambiguously identified. We used Chinese hamster ovary cells stably expressing 5-HT2C receptors as biodetectors to monitor 5-HT release from taste buds. When taste buds were depolarized with KCl or stimulated with bitter, sweet, or sour (acid) tastants, serotonin was released. KCl- and acid-induced 5-HT release, but not release attributable to sweet or bitter stimulation, required Ca2+ influx. In contrast, 5-HT release evoked by sweet and bitter stimulation seemed to be triggered by intracellular Ca2+ release. These experiments strongly implicate serotonin as a taste bud neurotransmitter and reveal unexpected transmitter release mechanisms.

Adenosine Triphosphate↗