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Lectin histochemistry on mucous substances of the taste buds and adjacent epithelia of different vertebrates.

In the present study carbohydrate residues in taste buds (TBs) and adjacent epithelial formations of a teleostean fish, a frog and the rabbit were detected by means of lectin histochemistry. Biotinylated lectins from Pisum sativum (PSA), Arachis hypogaea (PNA), Dolichos biflorus (DBA), Triticum vulgaris (WGA and succinylated WGA), Glycine max (SBA) and Ulex europaeus (UEA I) have been applied. The lectins were bound to an avidin-biotin-peroxidase-complex (ABC) and visualized by diaminobenzidine/H2O2. Most intensive reactivity was observed at the taste disc cells of the frog with DBA, S-WGA and SBA. PNA did not bind to the TBs of any of the animals tested. As shown in SBA preparations, sialic acid is present in a nonacylated and an acylated form in the mucosa of the frog's tongue. The TBs of the fish possess all the sugars we looked for except for the disaccharide D-galactose-(1-3)-beta-D-N-acetyl-galactosamine (Gal/GalNAc) and sialic acid. The TBs of the rabbit contain GalNAc, as detected with DBA, but not with SBA; and fucose (Fuc), mannose (Man) and N-acetyl-glucosamine (GlcNAc). As revealed by preincubation of the tissue sections with neuraminidase in TB cells of the rabbit, sialic acid masks Gal/GalNAc and GalNAc. These lectin-binding characteristics show that in the TBs of some selected representatives which belong to different vertebrate classes exist different mucous substances. These substances possess different binding characteristics to specific sugars, and this is possibly of particular interest to chemoreception phenomena.

Animals↗

Effects of genetic background on prostate and taste bud carcinogenesis due to SV40 T antigen expression under probasin gene promoter control.

The incidence of prostate carcinomas in African-American men is greater than in white men, indicating genetic factors are involved in risk of this neoplasia. Recently, we have developed a transgenic rat model of prostate cancer, featuring development of malignancies within 15 weeks of age at very high incidence. Male transgenic rats with a Sprague-Dawley genetic background were mated with wild-type females of F344, Wistar and ACI strains. F1 male transgenic hybrids with female Wistar and ACI rats had significantly lowered incidences of prostate carcinomas. However, the serum level of testosterone, and expression of the transgene, probasin, and the androgen receptor did not correlate with the strain variation in tumor development. Furthermore, immunohistochemical analysis of the SV40 Tag and the androgen receptor also did not reveal any differences between the strains. The transgenic rats additionally developed taste bud neuroblastomas at 100% incidence and this was suppressed in F1 male transgenic offspring with the ACI, but not the other strains. These results clearly show that genetic background influences prostate carcinogenesis and taste bud tumorigenesis in rats and that the present transgenic rats could provide a good model to identify specific factors.

Androgen-Binding Protein↗

The anatomical relations of the ductus vomeronasalis and the occurrence of taste buds in the papilla palatina of Nycticebus coucang (primates, prosimiae) with remarks on strepsirrhinism.

Nycticebus coucang is typically strepsirrhine. This condition is basically the same in all prosimians except Tarsius. The rhinarium shows a labial part; the marked furrow in its median plane clefts the margin of the upper lip. The latter is attached to the gum between the median incisors by means of paired folds (not corresponding to a frenulum labii superioris). The labial cleft is continued in the sulcus papillae palatinae. The ductus vomeronasalis opens into the ductus nasoplatinus which, in turn, enters the aboral part of the sulcus palatinus. Thus strepsirrhinism provides, occasionally, an open communication of the olfactory systems (nose, vomeronasal organ) with the environment in front of the rhinarium. Strepsirrhine primates possess an internarium very similar to the one of platyrrhine monkeys. The shape of the nostrils is not characteristics for strepsirrhinism. A split-lip condition is likely ancestral to the complete lip condition, since it is found in several stems of mammals, especially in archaic forms. Strepsirrhinism, such as in lemuroid prosimians, is probably just one case of the primitive split-lip condition. Therefore, a median furrow in the external hairy skin of the upper lip, found in some platyrrhine monkeys, could be a rudiment of a cleft, indicating that a kind of a split lip condition was ancestral to the undivided lips of platyrrhine monkeys. Taste buds occur in the epithelium of the lateral surface of the papilla palatina of Nycticebus. Other lorisid prosimians have not been studied in this respect. The taste buds test water soluble substances entering the sulcus either way. These substances may come from the environment entering the sulcus through the labial cleft.

Animals↗

Evidence that taste buds are not present on the human adult uvula.

We conducted a study to investigate whether taste buds are present on the human adult uvula. Our impetus was to determine whether surgical procedures that involve removal of the uvula can affect taste perception. Five human uvulae were removed via a modified carbon dioxide laser-assisted uvulopalatoplasty in an outpatient office setting. The uvulae were serially sectioned and stained with a solution specific for membrane-bound calcium-modulated adenosine triphosphatase, a high concentration of which is found in taste receptors. Examination of the stained sections under light microscopy failed to show that any taste receptors were present in any of the uvulae. This finding suggests that the taste disturbances noted after surgical procedures involving removal of the uvula are not attributable to a loss of taste receptors.

Adult↗

[Effect of substance P on the monoamine-containing cells of the taste buds].

The effect of substance P (SP) on monoamine-containing cells of the frog taste buds was studied by fluorescent microscopy. Intraperitoneal injection of SP resulted in a gradual increase of cell serotonin content. In monoamine deficiency caused by previous injection of rausedyl, SP favoured the recovery of the serotonin level to initial. When SP and rausedyl were used combined, SP protected the serotonin-containing cells from the depleting effect of rausedyl. The functional role of SP in the taste apparatus is discussed.

Animals↗

Lectin cytochemistry of the dark granules in the type 1 cells of Syrian hamster circumvallate taste buds.

Lectin-gold complexes in the dark granules (DGs) and the dense substance (DS) of vallate taste buds were localized. Both the DGs and the DS were labelled with wheat-germ agglutinin, Ulex europeus agglutinin-I and peanut agglutinin. Their common reaction to these lectins suggested that the DGs contain carbohydrate components similar to those of the DS. The results provide cytochemical evidence that the DS in the taste pit represents, at least in part, the content of the DGs in the type 1 cells.

Animals↗

Effects of extracellular pH, PCO2, and HCO3- on intracellular pH in isolated rat taste buds.

We studied the effects of changing external pH (pHo), external bicarbonate concentration ([HCO3-]o), and PCO2 on taste receptor cell (TRC) intracellular pH (pHi) in taste bud fragments (TBFs) isolated from rat circumvallate and fungiform papillae with the pH-sensitive fluoroprobe 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) using microfluorometric and imaging techniques. In N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid-buffered solutions, TRC pHi responded rapidly and monotonically to changes in pHo between 6.5 and 8.0. The relationship between pHi and pHo was steep, with slopes varying between 0.8 and 1.2. Similarly, varying pHo by changing PCO2 at constant [HCO3-]o or changing [HCO3-]o at constant PCO2 led to rapid, monotonic changes in pHi. The relationship between pHi and pHo was once again steep, with slopes varying between 0.8 and 1.2. However, simultaneous changes in PCO2 and [HCO3-]o at constant pHo did not cause any significant changes in steady-state pHi. In imaging studies, single, isolated TRCs responded to changes in pHo, with parallel changes in pHi in the soma and apical process. In addition, changes in pHo induced parallel changes in pHi throughout TBFs. These data suggest that the steady-state TRC pHi is a function of pHo. Changes in TRC pHi may be involved in acid sensing, and salivary [HCO3-] may play a role in the maintainance of steady-state TRC pHi and in the neutralization of acid-induced changes in pHi.

Animals↗

Membrane-bounded intratubular bodies in rat taste buds.

The ultrastructural features of membrane-bounded bodies contained in the tubulo-vesicular system in the outer segment of taste bud cells are described. Each body showed a round, fusiform or oval shape, was surrounded by a trilaminar membrane and enclosed an electron dense matrix sometimes containing inclusions. These bodies were found at all ages studied. Similar structures were also found embedded in the material plugging the taste pore. Our findings suggest that these bodies could be secreted at the free surface of the cells and be involved in the concentration of divalent cations.

Animals↗

[Possible nature of the biogenic amine in the dumbbell-shaped cells of frog taste buds].

An attempt was made to identify specific monoamines contained in the dumb-bell shape cells of the frog taste bud by means of histochemical analysis. It was shown by fluorescent microscopy that preliminary administration of exogenous serotonin into the blood channel of frog tongue resulted in a sharp increase of specific fluorescence of the dumb-bell shape cells, whereas serotonin synthesis inhibition with p-chlorphenylalanine led to reduction and elimination of specific fluorescence. It was concluded that-specific monoamine of the dumb-bell shape cells was possibly of serotonin-like nature.

Animals↗

[Scanning electron microscope study of the ontogeny of human lingual taste buds].

The surface of the tongue in the newborn and elderly subjects was examined by scanning microscopy to demonstrate some types of taste papilla. The lingual chemosensory auxiliary apparatus was shown to undergo heterochronic maturation in different periods of ontogenesis. The taste buds first matured in the papilla localized on the body and root and then on the apex of the tongue. The changes occurring in the chemosensory apparatus have been demonstrated for two age periods (for 60- and 80- year-old subjects).

Age Factors↗

[Pharmaco-histochemical study of monoamine-containing cells in the taste buds as affected by capsaicin-induced substance P deficiency].

A clearly positive correlation between the level of substance P (SP) in the taste nerves and cellular serotonin content in the taste buds has been established using fluorescent-histochemical technique. In chronic SP deficiency caused by capsaicin, cellular serotonin content was considerably decreased due to the disturbances in its synthesis and uptake. The injection of exogenous SP resulted in the recovery of the initial serotonin level. The data obtained confirm an important role of SP in the maintenance of the optimal cellular serotonin level owing to the activation of monoamine synthesis and uptake.

Animals↗

Noninvasive recording of receptor cell action potentials and sustained currents from single taste buds maintained in the tongue: the response to mucosal NaCl and amiloride.

Apical membrane currents were recorded from the taste pore of single taste buds maintained in the tongue of the rat, using a novel approach. Under a dissection microscope, the 150-microns opening of a saline-filled glass pipette was positioned onto single fungiform papillae, while the mucosal surface outside the pipette was kept dry. Electrical responses of receptor cells to chemical stimuli, delivered from the pipette, were recorded through the pipette while the cells remained undamaged in their natural environment. We observed monophasic transient currents of 10-msec duration and 10-100 pA amplitude, apparently driven by action potentials arising spontaneously in the receptor cells. When perfusing the pipette with a solution of increased Na but unchanged Cl concentration, a stationary inward current (from pipette to taste cell) of 50-900 pA developed and the collective spike rate of the receptor cells increased. At a mucosal Na concentration of 250 mM, the maximal collective spike rate of a bud was in the range of 6-10 sec-1. In a phasic/tonic response, the high initial rate was followed by an adaptive decrease to 0.5-2 sec-1. Buds of pure phasic response were also observed. Amiloride (30 microM) present in the pipette solution reversibly and completely blocked the increase in spike rate induced by mucosal Na. Amiloride also decreased reversibly the stationary current which depended on the presence of mucosal Na (inhibition constant near 1 microM). During washout of amiloride, spike amplitudes were first small, then increased, but always remained smaller than the amiloride-blockable stationary current of the bud.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗

Calbindin-like immunoreactivity in two peripheral chemosensory tissues of the rat: taste buds and the vomeronasal organ.

In the rat, calbindin-like immunoreactivity was observed at both the light and electron microscopic levels within the chemoreceptor neurons of the vomeronasal organ (VNO) and both intragemmal cells and associated nerve fibers of the circumvallate taste buds. All VNO neurons were immunoreactive. Only a subset of intragemmal taste cells was immunoreactive; associated immunoreactive nerve fibers were apposed to both labeled and unlabeled cells but no synaptic contacts were observed.

Animals↗

Immunocytochemistry on the localization of 5-hydroxytryptamine in monkey and rabbit taste buds.

Immuno-electron microscopy with the protein A-gold method demonstrated immunoreactive gold particles against 5-hydroxytryptamine localized in cored vesicles aggregating around presynaptic terminals of the gustatory cells in monkey and rabbit taste buds. The positive reactions were also found in the intragemmal and subepithelial nerve fibers. The role of these cored vesicles in taste transduction is still uncertain but the data strongly suggest that they may participate in a serotonergic modulation of a cholinergic synaptic transmission from the gustatory cells to the nerve endings.

Animals↗

Taste bud form and distribution on lips and in the oropharyngeal cavity of cardinal fish species (Apogonidae, Teleostei), with remarks on their dentition.

The oral dentition and type and number of taste buds (TB) on the lips and in the oropharyngeal cavity were compared by means of SEM in 11 species of cardinal fishes (Apogonidae) belonging to five genera. The occurrence of a dense cover of skin papillae on the lips of some species (e.g., Apogon frenatus), as well as differences in structure of vomer, tongue, and palatinum, expose additional morphological characters important for clarification of the taxonomy of this group of fishes. Differences are also revealed in the type of dentition, such as on the vomer and epi-hypopharyngeal bones. Strong and dense dentition of the anterior part of the oral cavity and a high number of TB on this site in species feeding on larger prey (e.g., Cheilodipterus spp) is compared to the relatively feeble jaw armor and richness of TB on the more pharyngeal site in species feeding on smaller prey (e.g., Apogon angustatus, A. frenatus). In addition to the three types of TB (Types I-III) previously described from various teleost fish, a fourth type (Type IV), comprising very small buds, was found in some cardinal fish (Apogon angustatus, A. frenatus). The various TB are distributed from the lips to the pharyngeal bones, on the breathing valves, tongue, palatinum, and pharyngeal bones; their number and type on the various sites differ in the different species. In all species studied the Types I and II TB, elevated above the surrounding epithelium, dominated the lips and anterior part of mouth, while Types III and IV, which end apically at the level with the epithelium, dominated the more posterior pharyngeal region. The highest number of TB, around 24,600, were found in Fowleria variegata, a typical nocturnal species, and the lowest in the diurnal and crepuscular Apogon cyanosoma (1,660) and Cheilodipterus quinquestriatus (2,400). Differences are also revealed in the type of dentition, such as on the vomer and epi-hypopharyngeal bones. The number of TB increased with growth of the fishes. The differences in the total number of TB and their distribution in the oropharyngeal cavity in the various species indicates possible different mechanisms of foraging and food-recognition.

Animals↗

Synaptobrevin-2-like immunoreactivity is associated with vesicles at synapses in rat circumvallate taste buds.

Synaptobrevin is a vesicle-associated membrane protein (VAMP) that is believed to play a critical role with presynaptic membrane proteins (SNAP-25 and syntaxin) during regulated synaptic vesicle docking and exocytosis of neurotransmitter at the central nervous system. Synaptic contacts between taste cells and nerve processes have been found to exist, but little is known about synaptic vesicle docking and neurotransmitter release at taste cell synapses. Previously we demonstrated that immunoreactivity to SNAP-25 is present in taste cells with synapses. Our present results show that synaptobrevin-2-like immunoreactivity (-LIR) is present in approximately 35% of the taste cells in rat circumvallate taste buds. Synaptobrevin-2-LIR colocalizes with SNAP-25-, serotonin-, and protein gene product 9.5-LIR. Synaptobrevin-2-LIR also colocalizes with immunoreactivity for type III inositol 1,4,5-triphosphate receptor (IP3R3), a taste-signaling molecule in taste cells. All IP3R3-LIR taste cells express synaptobrevin-2-LIR. However, approximately 27% of the synaptobrevin-2-LIR taste cells do not display IP3R3-LIR. We believe, based on ultrastructural and biochemical features, that both type II and type III taste cells display synaptobrevin-2-LIR. All of the synapses that we observed from taste cells onto nerve processes express synaptobrevin-2-LIR, as well as some taste cells without synapses. By using colloidal gold immunoelectron microscopy, we found that synaptobrevin-2-LIR is associated with synaptic vesicles at rat taste cell synapses. The results of this study suggest that soluble NSF attachment receptor (SNARE) machinery may control synaptic vesicle fusion and exocytosis at taste cell synapses.

Animals↗