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Expression and physiological actions of cholecystokinin in rat taste receptor cells.

Gustatory perception arises not only from intracellular transduction cascades within taste receptor cells but also from cell-to-cell communication among the cells of the taste bud. This study presents novel data demonstrating that the brain-gut peptide cholecystokinin (CCK) is expressed in subsets of taste receptor cells, and that it may play a signaling role unknown previously within the taste bud. Immunocytochemistry revealed positively stained subsets of cells within taste buds throughout the oral cavity. These cells typically displayed round nuclei with full processes, similar to those classified as light cells. Peptide expression was verified using nested PCR on template cDNA derived from mRNA extracted from isolated posterior taste buds. Multiple physiological actions of cholecystokinin on taste receptor cells were observed. An outward potassium current, recorded with the patch-clamp technique, was inhibited by exogenous application of sulfated cholecystokinin octapeptide in a reversible and concentration-dependent manner. Pharmacological analysis suggests that this inhibition is mediated by CCK-A receptors and involves PKC phosphorylation. An inwardly rectifying potassium current, typically invariant to stimulation, was also inhibited by cholecystokinin. Additionally, exogenous cholecystokinin was effective in elevating intracellular calcium as measured by ratiometric techniques with the calcium-sensitive dye fura-2. Pharmacology similarly demonstrated that these calcium elevations were mediated by CCK-A receptors and were dependent on intracellular calcium stores. Collectively, these observations suggest a newly discovered role for peptide neuromodulation in the peripheral processing of taste information.

Animals↗

Role of the G-protein subunit alpha-gustducin in taste cell responses to bitter stimuli.

Many bitter stimuli are believed to bind to specific G-protein-coupled membrane receptors on taste cells. Despite the compelling evidence for its pivotal role in bitter taste sensation, a direct involvement of the G-protein subunit alpha-gustducin in bitter taste transduction in taste cells has not been demonstrated in situ at the cellular level. We recorded activation of taste cells by bitter stimuli using Ca2+ imaging in lingual slices and examined alpha-gustducin immunoreactivity in the same cells. In mice vallate papillae, many, but not all, bitter-responsive cells expressed alpha-gustducin. In agreement with this correlation, the incidence of cells responding to bitter stimuli was reduced by 70% in mutant mice lacking alpha-gustducin. Nevertheless, some taste cells lacking alpha-gustducin responded to bitter stimuli, suggesting that other G-protein alpha subunits are involved. We found that the G-protein alpha subunit Galpha(i2) is present in most bitter-responsive cells and thus may also play a role in bitter taste transduction. The reduced behavioral sensitivity to bitter stimuli in alpha-gustducin knock-out mice thus appears to be the consequence of a reduced number of bitter-activated taste cells, as well as reduced sensitivity.

Animals↗

Proliferation of taste buds in the foliate and vallate papillae of postnatal hamsters.

The growth of the taste system in the hamster is considered in comparison to the postnatal development of other organ systems and the entire animal. No taste buds are present in vallate or foliate papillae of the hamster at birth, but they attain both the appearance and numbers of adult taste buds within 5 weeks of age. The most rapid increase in the number of taste buds occurs within the first 10 days of life, and this proliferation anticipates the weaning of hamsters which occurs by about three weeks of age. Foliate taste buds reach a maximal number within two months, but vallate taste buds continue to increase in number through 4 months of age. Taste bud proliferation and development occur earlier and more rapidly than in other organ systems. This early development of taste buds may protect the weanling hamster against accidental poisoning by noxious plants, and it may also reinforce the food intake which is required for normal growth.

Animals↗

Taste threshold determinations on dyspeptic patients and on blind controls.

Taste threshold detection and recognition were determined on 46 dyspeptic patients and on 42 blind controls. Among the dyspeptic patients 60% suffered from different taste disorders. Among the blind controls the frequency of taste disorder was 41%. Among the dyspeptic patients heterogeusia and dysgeusia dominated, threefold compared to the control blind persons. A mild hypergeusia appeared among the patients in 21%, among the blind in 8%. There was an enhanced hypergeusia in the group of blind persons (16%), but this was found in the group of dyspeptic patients in only 4%. It is suggested that the hypergeusia may also be a compensatory function, a consequence of the loss of seeing. There was an inverse ratio between the taste acuity and taste preference. The taste threshold to the preferred taste in both groups was higher, therefore more food was needed to stimulate the taste nerves.

Adult↗

[Evaluating function and disorders of taste].

In a first anatomical section the peripheral gustatory pathways, their central connections, nuclei and cortical projections are discussed. It is evident, that the gustatory fibres from the posterior part of the tongue run in the IX nerve and those from the soft palate reach the medulla oblongata via the petrosal and facial nerve. For the anterior part of the tongue there obviously exists only one gustatory pathway via the chorda tympani-facial nerve. About the further central pathways of taste fibres is much less known. In a second part the methods of taste testing with different taste solutions and the electrogustometry are described. Their practical use and the pitfalls of testing are considered. The disorders of the taste sense compose a third part. Genetic and endocrine abnormalities as well as the side effects of drugs and radiotherapy and the destruction of taste nerves may lead to gustatory deviations. The possibility of central gustatory disorders, especially the combined loss of taste and smell as a result of trauma are mentioned. A chapter dealing with the therapy of taste disorders and a short outlook on the genetic aspects of this oral sense complete this review.

Afferent Pathways↗

Taste and smell losses in normal aging and disease.

OBJECTIVE: To review the scientific literature on the alterations in the senses of taste and smell in the elderly, including causes, diagnosis, prognosis, and treatment. DATA SOURCES: Original reports and reviews obtained through MEDLINE searches from 1966 through June 1997 using the MeSH headings of "taste," "taste buds," "taste disorders," "taste thresholds," "smell," "odors," "aged," and "aging." Articles frequently cited in reference lists were also included. STUDY SELECTION: All articles were reviewed, tabulated, and summarized. DATA EXTRACTION: Criteria for extraction included data quality and validity, statistical treatment of the data, venue of publication, and relevance to clinical care. CONCLUSION: Losses of taste and smell are common in the elderly and result from normal aging, certain disease states (especially Alzheimer disease), medications, surgical interventions, and environmental exposure. Deficits in these chemical senses cannot only reduce the pleasure and comfort from food, but represent risk factors for nutritional and immune deficiencies as well as adherence to specific dietary regimens. Chemosensory decrements can lead to food poisoning or overexposure to environmentally hazardous chemicals that are otherwise detectable by taste and smell. Use of flavor-enhanced food can increase enjoyment of food and have a positive effect on food intake and immune status.

Aged↗

Effects of stimulus preexposure on the generalization of conditioned taste aversions in infant rats.

Generalization of a conditioned taste aversion in infant rats and how this is affected by stimulus preexposure was investigated in a series of experiments. In Experiment 1 generalization of a conditioned aversion between two tastes (sweet and salty) was found, and the effect of tastes preexposure was a reduction in generalization (Experiment 2). However, when these tastes were combined with a common taste (acid) that was less (Experiment 3) or more intense (Experiment 3b), the effect of stimulus preexposure was a stronger generalization of the conditioned aversion. In this case, a reduction on generalization was again observed by increasing the number of preexposure trials to the taste compounds (Experiment 4). In all cases the generalization levels were directly related to the effect of stimulus preexposure on the acquisition rate of conditioning. It can be concluded that, with the appropriate parameters, a reduction of generalization of a conditioned taste aversion can be obtained after taste exposure in preweanling rats.

Animals↗

Taste cells with synapses in rat circumvallate papillae display SNAP-25-like immunoreactivity.

SNAP-25 is a 25 kDa protein believed to be involved in the processes of membrane fusion and exocytosis associated with neurotransmitter release. In the present study we present evidence that SNAP-25-like immunoreactivity can be used as a marker for taste cells with synapses in rat circumvallate papillae. SNAP-25 immunoreactivity is present in most intragemmal nerve processes and a small subset of taste cells. Intense immunoreactivity is associated with the nerve plexus located below the base of the taste bud. Of a total of 87 taste cells with synapses onto nerve processes, 80 of the presynaptic taste cells had SNAP-25 immunoreactivity. The association of SNAP-25 immunoreactivity with taste cells possessing synapses suggests that these cells may be gustatory receptor cells. Because this SNAP-25 antibody can label taste cells with synapses, it may also serve as a useful tool for future studies correlating structure with function in the taste bud.

Animals↗

Characterization of different microenvironments at the surface of the frog's taste organ.

We used a panel of histochemical techniques to identify and characterize the cell-associated extracellular material at the surface of the frog's taste organ. We employed morphological and histochemical techniques using both the light microscope and the electron microscope. Results show that the apical, external aspect of cells reaching the surface of the taste organ is in close contact with a layer of amorphous material. The histochemical characteristics of this material vary according to the cell type with which it is in contact. Three different microenvironments can be identified at the surface of the frog's taste organ: type 1 microenvironment is associated with the superficial layer of mucus (secretory) cells; type 2 microenvironment characterizes the surface of the so-called wing cells, which reach the surface of the taste organ as thin laminae running among mucus cells; and type 3 microenvironment shrouds the free endings of putative taste cells and is rich in calcium and lipids. Type 2 and type 3 microenvironments fix peroxidase (a sapid macromolecule) with increasing affinity. We conclude that highly differentiated microenvironments exist at the surface of the frog's taste organ, and these could play a role in the chain of biological events leading to the taste sensation. Furthermore, characterization of the cell-associated, specific microenvironments can help clarify the role of the different cell types in the frog's taste organ.

Animals↗

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

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

Animals↗

Group IIA phospholipase A(2) is coexpressed with SNAP-25 in mature taste receptor cells of rat circumvallate papillae.

The taste buds are composed of heterogeneous cell populations with diverse properties and at different stages of maturity. It is important to define the relationships between cell properties and cell maturity to understand the molecular events involved in intracellular taste signaling. In the present study, in situ hybridization analysis indicated that group IIA phospholipase A(2) (PLA(2)-IIA) is expressed in a subset of taste bud cells. Immunohistochemical studies showed that PLA(2)-IIA was expressed in a subset of cells expressing phospholipase Cbeta2, a molecule essential for taste signaling in taste receptor cells, and also that some PLA(2)-IIA-positive cells expressed gustducin (Ggust), a bitter-taste-signaling molecule. Although PLA(2)-IIA and Ggust were expressed at similar frequencies in taste buds, bromodeoxyuridine (BrdU) chase experiments indicated that the expression of Ggust began 2 days after BrdU injection, whereas the expression of PLA(2)-IIA commenced after 4 days. In addition, PLA(2)-IIA was coexpressed with SNAP-25, a synaptosomal-associated protein. These results indicated that PLA(2)-IIA is expressed in mature taste receptor cells that possess exocytotic machinery.

Animals↗

Nucleoside triphosphate diphosphohydrolase-2 is the ecto-ATPase of type I cells in taste buds.

The presence of one or more calcium-dependent ecto-ATPases (enzymes that hydrolyze extracellular 5'-triphosphates) in mammalian taste buds was first shown histochemically. Recent studies have established that dominant ecto-ATPases consist of enzymes now called nucleoside triphosphate diphosphohydrolases (NTPDases). Massively parallel signature sequencing (MPSS) from murine taste epithelium provided molecular evidence suggesting that NTPDase2 is the most likely member present in mouse taste papillae. Immunocytochemical and enzyme histochemical staining verified the presence of NTPDase2 associated with plasma membranes in a large number of cells within all mouse taste buds. To determine which of the three taste cell types expresses this enzyme, double-label assays were performed with antisera directed against the glial glutamate/aspartate transporter (GLAST), the transduction pathway proteins phospholipase Cbeta2 (PLCbeta2) or the G-protein subunit alpha-gustducin, and serotonin (5HT) as markers of type I, II, and III taste cells, respectively. Analysis of the double-labeled sections indicates that NTPDase2 immunoreactivity is found on cell processes that often envelop other taste cells, reminiscent of type I cells. In agreement with this observation, NTPDase2 was located to the same membrane as GLAST, indicating that this enzyme is present in type I cells. The presence of ecto-ATPase in taste buds likely reflects the importance of ATP as an intercellular signaling molecule in this system.

Adenosine Triphosphatases↗

Regeneration of fungiform taste buds: temporal and spatial characteristics.

The gross morphology of the tongue of the Mongolian gerbil Meriones unguiculatus), the location of papillae and taste buds, and the normal innervation pattern of the tongue and taste buds were determined. The chorda tympani nerve was interrupted to produce degeneration of fungiform taste buds. Regenerating chorda tympani axons followed the original nerve pathways in the tongue en route to the fungiform papillae in the epithelium where they initiated the regeneration of taste buds. The spatial distribution of reinnervated fungiform papillae and reformed taste buds was examined 7 to 19 days following surgery. Beginning at eight days following chorda tympani interruption there was a progressive increase, first, in the proportion of fungiform papillae that were reinnervated, and later in the number of reformed taste buds. On the basis of these measures it was concluded that a taste bud is reformed one to two days after reinnervation of its papilla. From the time course of reinnervation of the fungiform papillae it was calculated that some fibers regenerated at rates in excess of 2 mm/day. Regeneration was precise and systematic. The regenerating chorda tympani fibers accurately returned to the fungiform papillae; they did not follow the pathways of lingual nerve axons. In the initial stages of recovery both reinnervated papillae and reformed taste buds were preferentially located toward the front of the tongue; the reinnervation of posterior fungiform papillae was delayed.

Animals↗

Ultrastructure of taste cells and synapses in the mudpuppy Necturus maculosus.

Taste buds in the mudpuppy Necturus maculosus were examined with electron microscopy. Three cell types (dark, light, and basal) were identified and reconstructed from serial thick sections. Dark and light cells extend from the basal lamina to the surface of the tongue. The apical process of the dark cells was usually quite lamellar when viewed in cross section, in contrast to light cells, whose apical process appeared more cylindrical. Basal cells are situated at the base of the bud and do not extend processes to the surface of the tongue. The cytoplasm of basal cells contains numerous clear and dense-cored vesicles. Small, spinelike processes (2-3 microns in length) project outward from the basal cells into the cytoplasm of the surrounding tast receptor cells. Morphologically, basal cells in mudpuppy taste buds resemble Merkel cells. Unmyelinated afferent nerve fibers enter the taste bud at the base and course through the lower portion of the bud. Synapses were found between taste receptor cells and nerve fibers, between basal cells and nerve fibers, and between basal cells and taste receptor cells. Over 65% of the synapses observed in the mudpuppy taste bud involved the basal cell. These findings suggest that basal cells play some role in chemosensory signal processing or integration of the taste response.

Animals↗

Development of fungiform papillae, taste buds, and their innervation in the hamster.

Fungiform taste buds in mature hamsters are less subject to neurotrophic influences than those of other species. This study evaluates taste-bud neurotrophism during development in hamsters by examining the relation between growing nerves and differentiating fungiform papillae. Chorda tympani (CT) or lingual (trigeminal) nerve (LN) fibers were labelled with Lucifer Yellow as they grew into (CT fibers) or around (LN fibers) developing taste buds. Developing fungiform papillae and taste pores were counted with the aid of a topical tongue stain. The tongue forms on embryonic days (E) 10.5-11 and contains deeply placed CT and LN fibers but no papillae. By E12, the tongue epithelium develops scattered elevations. These "eminences" selectively become innervated by LN fibers that grow to the epithelium earlier and in larger numbers than CT fibers. Definitive fungiform papillae form rapidly during E13-14 and become heavily innervated by LN fibers. Intraepithelial CT fibers, rare at E13, invariably innervate fungiform papillae containing nascent taste buds at E14. During E14-15 (birth = E15-16), most papillae contain taste buds with pores, extensive perigemmal LN innervation, and extensive intragemmal CT innervation. At birth, numbers of fungiform papillae and taste pores are adultlike. The results show that fungiform eminences begin forming in the absence of innervation. The subsequent differentiation of definitive fungiform papillae and their innervation by LN fibers occur synchronously, prior to the differentiation of taste buds and their CT innervation. The hamster is precocious (e.g., compared to rat) in terms of LN development and the structural maturity of the anterior tongue at birth.

Animals↗

Cell contact-dependent mechanisms specify taste bud pattern during a critical period early in embryonic development.

After gastrulation, the pharyngeal endoderm is specified to give rise to taste receptor organs without further signaling from other embryonic tissues. We hypothesized that intercellular signaling might be responsible for the specification of taste buds. To test if and when this signaling was occurring, intercellular contacts were transiently disrupted in cultures of pharyngeal endoderm from axolotl embryos, and the number, size, and distribution of taste buds analyzed. Disruption of cell contacts at progressive time points, from neurula to late tail bud stages, revealed a critical period, during mid-tail bud stages, when disruption of cell contacts resulted in a significant increase in taste bud number and size. The spatial distribution of taste buds was also altered; taste buds were more clustered in explants disrupted during the critical period. These effects were not due to general alterations in mitosis and apoptosis. Rather, at least three aspects of taste bud patterning, i.e., number, size, and distribution, are governed by mechanisms dependent on normal cell contacts during a concise time window. Furthermore, our findings are consistent with specification of taste buds by means of lateral inhibitory signaling, which we hypothesize results from cell contact-dependent or short-range diffusible signals.

Ambystoma↗

Effects of prior exposure to palatable and unpalatable novel foods on children's willingness to taste other novel foods.

In two studies, 7- to 9-year-old and 10- to 12-year-old children received taste exposure to four good-tasting familiar, four good-tasting novel or four bad-tasting novel foods. Following this exposure phase, they saw a series of different foods, familiar and novel, and rated their willingness to taste them. For older children, exposure to the novel-good foods increased willingness to taste novel foods in comparison to the familiar-good control, while exposure to the novel-bad foods had no effect. For younger children, exposure to both novel-good and novel-bad foods decreased willingness to taste novel foods. The studies were originally framed in terms of children's schemas about novel foods and how exposure to good- and bad-tasting novel foods constituted provision of schema-inconsistent (novel-good) or schema-consistent (novel-bad) information. While such a framework accounted well for the results for the older children, it did not account for those for the younger children. The behaviour of the younger children was tentatively explained in terms of their attempt to regulate arousal produced by the initial exposure to the novel foods.

Child↗

The effect of flavor principles on willingness to taste novel foods.

E. Rozin and P. Rozin have suggested that one of the functions of "flavor principles" (the distinctive seasoning combinations which characterize many cuisines) is to facilitate the introduction of novel staple foods into a culture by adding sufficient familiarity to decrease the neophobia ordinarily produced by a new food. We tested this idea experimentally, predicting that the addition of a familiar flavor principle to a novel food would increase individuals>> willingness to taste it, in comparison to their willingness to taste the same food in the absence of the flavor principle. Since people have little reluctance to approach familiar foods, addition of a familiar flavor principle to a familiar food should have little effect on willingness to taste it. In a pilot study, subjects selected from a list, a sauce which was high in familiarity and liking for them. They then rated their willingness to taste one novel and one familiar food with the sauce and one of each with no sauce. Subjects did not actually see any foods-the familiar and novel foods were simply described-and they were aware that they would not actually be tasting any foods. In the study proper, subjects rated their willingness to try each of the four food/sauce combinations described above. In this study, they actually saw the foods they were rating and were under the impression their ratings would determine what they would taste later in the study. In both studies, the addition of a familiar sauce to a novel food increased subjects>> willingness to taste it (in comparison to the same food with no sauce) while the addition of a sauce to a familiar food either had no effect or decreased subjects>> willingness to taste it. In the pilot study, the "flavor principle" effect interacted with subjects>> levels of food adventurousness.

Adolescent↗