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Taste reactivity to alcohol and basic tastes in outbred mice.

The taste reactivity test was used to determine the response of outbred mice to orally infused taste solutions. For the initial measures, mice (n = 10) were tested with 3%, 6%, 9%, and 12% (v/v) alcohol and four taste solutions: sucrose, sodium chloride, hydrochloric acid, and quinine hydrochloride (a single concentration of each). A second group of naive mice (n = 16) was tested with 5%, 10%, 20%, 30%, and 40% alcohol. The final set of measures with naive mice (n = 26) was taken with a range of sucrose concentrations: 0.01 M, 0.05 M, 0.1 M, 0.5 M, and 1.0 M. In general, mice made similar reactivity responses to all solutions tested. A predominant component of the mouse response to all infused fluids was forelimb flailing; gaping was also a common response to all solutions. Despite the large number of aversive-type responses, mice rejected very little fluid via passive drip or fluid expulsion. The single, significant difference in responding to the four taste stimuli was that mice made fewer aversive responses to sucrose. Differential responding to the 5 to 40% alcohol concentrations and sucrose concentrations was observed. Mice increased ingestive responding as the concentration of alcohol and sucrose increased. Aversive responding decreased reliably only with increases in the sucrose concentration. Data provide the first reported taste reactivity responses of mice to orally infused taste solutions. These results can be compared with the extant data available in rats and can also be used as a basis for exploring taste factors in genetically defined mouse populations.

Alcohol Drinking↗

Bovine circumvallate taste buds: taste cell structure and immunoreactivity to alpha-gustducin.

The taste buds of bovine circumvallate papillae were investigated under light and electron microscopy both by histological and immunohistochemical methods. Taste buds existed in the inner epithelium of the trench of the papillae. Under electron microscopy, two types of taste cells, type I and type II, could be classified according to the existence of dense-cored vesicles and cytoplasmic density. Type I had electron-lucent cytoplasm and possessed many electron-dense cored vesicles in the apical cytoplasm. It was considered that the electron-dense materials of the vesicles were released and constituted the pore substance. This type of cell possessed long and thick apical processes in the taste pore. Type II had denser electron cytoplasm compared with that of type I and possessed many electron-lucent vesicles in the apical cytoplasm. This type of cell possessed microvilli in the taste pore. To know the immunoreactivity to alpha-gustducin in bovine circumvallate taste buds, we used the immunoblotting method and the immunohistochemical method. The alpha-gustducin reaction band at 40 kDa was displayed in the specimen of Western blots. The immunohistochemical property of the antiserum to alpha-gustducin was investigated by using the avidin-biotin complex (ABC) method and the 1.4-nm gold and silver enhancement methods. A subset of taste cells showed the immunoreactivity under light microscopy. The electron microscopic specimens with the 1.4-nm gold and silver enhancement method revealed that only type II cells exhibited the alpha-gustducin immunoreactivity.

Animals↗

Ultrastructural evidence for a binding substance to the sweet-tasting protein thaumatin inside taste bud pores of rhesus monkey foliate papillae.

Thaumatin is a protein that tastes intensely sweet only to Old World monkeys and to higher primates, including man. Here we used pre-embedding ultrastructural methods to study the distribution of thaumatin in apical regions of Rhesus monkey foliate papillae, using thaumatin conjugated to 5 nm gold particles. With freeze-substitution we saw that gold-labeled thaumatin bound to an electron-opaque, sponge-like secretory substance inside the taste bud pores. Labeled thaumatin was found at the surface of the secretory substance even deep inside the pore, where other, unlabeled cellular structures surrounded the substance. With freeze-fracture deep-etching the secretory substance that bound the thaumatin-gold particles appeared coarsely granular. There was no labeling of any other taste bud pore structure, including microvilli and small membrane-lined vesicles. Pre-incubation with an excess of unlabeled thaumatin inhibited binding with gold-labeled thaumatin. The results suggest that the secretory substance had the greatest affinity of all taste pore structures to the sweet-tasting compound under our experimental conditions. Therefore, gustatory reception probably involves various taste compound binding structures, microvilli, and also secretory substances like the one described here which bound thaumatin. We speculate that the secretory substance may bind taste stimuli and serve as an intermediate between stimuli and receptors. It could be involved in stimulus removal or delivery or both.

Animals↗

The ultrastructure of taste and touch receptors of the frog's taste organ.

The taste buds from fungiform papillae and the hard palate of frogs were investigated with the scanning and transmission electron microscopes. An immature pre-taste cell and a mature taste cell can be differentiated. Only the mature taste cell exhibits synaptic contact with the afferent taste fibre. Glandular and satellite supporting cells envelop the thin apical processes of the sensory cells. At the base of the taste disc up to 10 Merkel cells form a complex with nerve endings. There are two types of myelinated fibres, large and small. The small fibre innervates the taste cells, the thicker nerve fibre the Merkel cells. The occurrence of two types of receptors explains physiological results.

Animals↗

Location of taste buds in intact taste papillae by a selective staining method.

Taste buds were found to stain strongly and selectively in intact papillae with highly acidic dyes such as ponceau S. In intact tongues the taste buds in the fungiform, circumvallate and foliate papillae of the cynomolgus monkey and in the fungiform papillae of the rat as well as the taste discs in the fungiform papillae of the frog could be visualized. This method enables a rapid location and counting of taste buds in taste papillae without preparing histological sections. In cynomolgus tongue material fixed in formalin, the dyes penetrate into the buds. In fresh tongues only the taste pore region of the buds stains, which suggests that in vivo taste buds are impenetrable underneath the pore.

Animals↗

Lipid profiles of taste and non-taste epithelial tissues from steer tongues.

Some hypotheses on taste reception have implicated lipids of taste cells as major receptor constituents. This study reports detailed lipid profiles of the taste bud-containing epidermis from circumvallate papillae and fungiform papillae as well as profiles from two non-taste bud tissues: circumvallate papillae dermis and epidermis from the lateral posterior of the tongue. Differences in levels of triglycerides and phosphatidylcholines were observed but these were not directly related to the presence of taste buds. At this level of analysis, it is evident that there are no unusual distributions of phospholipid classes in the taste bud epidermis when compared with the non-taste bud lingual epidermis.

Animals↗

Taste responses to electrolytes in the frog glossopharyngeal nerve: initial process of taste reception.

In taste reception, it has been proposed that changes in surface potential on the apical membrane of taste cells bring about activation of taste cells during chemical stimulation. To ascertain whether changes in the surface potential are involved in taste reception of electrolytes, unitary discharges were recorded from single water fibers of the frog glossopharyngeal nerve with a suction electrode. The surface potential is a function of both the charge density of the membrane surface and the ionic strength of the medium. Low concentrations of CaCl2 (less than 1 mM) were very effective stimuli. However, 0.01-1 mM LaCl3 and HCl (pH 3.0-4.5), which alter the surface potential in the positive direction, had no excitatory effect. Transition metal cations, such as Mn2+, Co2 and Ni2+, had an excitatory effect, but the responses to these cations appeared at relatively high concentrations (greater than 5 mM), in spite of the high affinity of the receptor membrane for these cations. The results suggest that the surface charge of the apical membrane is not associated with the excitation caused by electrolytes. MgCl2 (greater than 5 mM) and NaCl (greater than 100 mM) were also effective stimuli, whereas choline Cl (100-1000 mM) had no excitatory effect. An increase in the ionic strength was achieved by the addition of 100-300 mM choline Cl to stimulating solutions of MgCl2 or NaCl. The responses to Mg2+ and Na+ were not affected by the increase in the ionic strength. The results obtained here indicate that changes in the surface potential on the surface of the apical membrane are not involved in taste reception of electrolytes. Alteration of the surface potential of the membrane in the positive direction would bring about a reduction in the local concentration of cations in the vicinity of the membrane. Hence, the presence of divalent cations in the medium may affect the response to monovalent cations. However, addition of 100 mM MgCl2 to the stimulating solution of NaCl did not affect the concentration-response curve for NaCl. This result suggests that the surface charge density of the apical membrane is very low and hence the magnitude of the surface potential is very small. The results also suggest that Mg2+ and Na+ activate the taste cells by two separate, non-interacting processes. The present study suggests that, in the initial process of taste reception, only the binding of each separate cation to its appropriate receptor site (specific receptor site) leads to activation of the receptor.

Animals↗

Transient impairment of cholinergic function in the rat insular cortex disrupts the encoding of taste in conditioned taste aversion.

The muscarinic antagonist scopolamine blocks conditioned taste aversion (CTA) when microinjected bilaterally into the rat insular cortex shortly before the exposure of the rat to a novel taste (the conditioned stimulus, CS) in CTA training. Scopolamine has no effect when microinjected shortly after the exposure to the novel taste or shortly before the application of the malaise-inducing agent (unconditioned stimulus, UCS). Scopolamine does not affect sensory, motor and retrieval mechanisms required for performing the CTA task, and does not block CTA when injected into another cortical area. The effect of scopolamine is independent of the taste used as CS. Furthermore, microinjection of scopolamine into the insular cortex shortly before the pre-exposure to a new taste in a latent inhibition paradigm, impairs the attenuation of CTA by that pre-exposure. Other muscarinic antagonists, pirenzepine and AF DX-116, have an effect similar to that of scopolamine. Comparison of the dose-dependency curves of the muscarinic antagonists suggests a predominant role in CTA for M2 subtype receptors. Carbachol, a muscarinic agonist, also impairs the encoding of taste in the insular cortex, but the results are confounded by the ability of that ligand to induce seizures. Our findings suggest that cholinergic neuromodulation participates in processing the CS in the gustatory cortex in CTA, either by encoding novelty at the cellular level, or by instructing the neural circuits to store the novel taste representation.

Animals↗

Heterogeneous distribution of taste cells in facial and vagal nerve-innervated taste buds.

Input from the three gustatory nerves of vertebrates is used to evaluate the nutritional quality of food. In some species, these cranial nerves are modified to accomplish additional specific functions. For example, the facial nerve innervated taste buds distributed over the body surface of catfish aid food search. Physiological studies indicate that this extra-oral taste pathway is more sensitive to amino acids than either the glossopharyngeal or vagal systems of the oral cavity. The current investigation seeks to determine if differences in taste cell subtypes might contribute to the observed differences in sensitivity. The distributions of five low molecular weight metabolites, L-alanine, L-aspartate, L-glutamate, GABA, taurine and the tripeptide glutathione, were examined in 2118 individual taste cells innervated by either the facial or vagal nerve of the channel catfish, Ictalurus punctatus. The metabolite profiles of these cells were determined immunocytochemically and subjected to a k-means clustering algorithm. Fifteen cell classes with quantitatively different patterns of metabolite co-localization were identified. All but one small class of two cells were found in both facial and vagal nerve-innervated taste buds. Four classes (9% of the total cells) had high, two classes (17%) had intermediate and the remaining nine classes (74%) had low levels of GABA immunoreactivity. While the functional significance of differences in metabolite profile remains to be determined, taste cell classes were not uniformly distributed across vagal and facial nerve innervated taste buds and may provide an anatomical basis for previously reported differences in gustatory sensitivity.

Algorithms↗

Polycose taste pre-exposure fails to influence behavioral and neural indices of taste novelty.

Taste novelty can strongly modulate the speed and efficacy of taste aversion learning. Novel sweet tastes enhance c-Fos-like immunoreactivity (FLI) in the central amygdala and insular cortex. The present studies examined whether this neural correlate of novelty extends to different taste types by measuring FLI signals after exposure to novel and familiar polysaccharide (Polycose) and salt (NaCl) tastes. Novel Polycose not only failed to elevate FLI expression in central amygdala and insular cortex, but also failed to induce stronger taste aversion learning than familiar Polycose. Novel NaCl, on the other hand, showed patterns of FLI activation and aversion learning similar to that of novel sweet tastes. Possible reasons for the resistance of Polycose to typical pre-exposure effects are discussed.

Analysis of Variance↗

Expression and localization of amiloride-sensitive sodium channel indicate a role for non-taste cells in taste perception.

Salty taste is blocked by the diuretic amiloride, which inhibits specific sodium channels. We have isolated an amiloride-sensitive sodium channel (ASSC) from taste tissues by polymerase chain reaction and screening of a cDNA library prepared from rat circumvallate papillae. Northern analysis reveals ASSC in taste and non-taste tissues with the highest level of expression of ASSC in the lung. In situ hybridization establishes ASSC localizations in the epithelia of lung and colon as well as tongue epithelial layers containing and lacking taste buds. These results support a model in which ASSC in non-taste cells regulates responses of taste cells to salt as well as other tastants.

Amiloride↗

A new taste reactivity analysis of the integration of taste and physiological state information.

We used conjoint manipulation of taste and physiological state to address the theoretical issue of signal integration. The interaction between taste (glucose concentration) and state (food deprivation) was evaluated using the taste reactivity method in which oral motor responses elicited by direct intraoral infusion are measured. The time frame of the typical taste reactivity paradigm, where observation is limited to the infusion period, was expanded to include the postinfusion interval. In each test session, rats received a series of trials consisting of 15-s intraoral infusions and 45-s postinfusion observation intervals. Two experiments were run in which glucose concentration was varied and rats were run nondeprived and after 24 h food deprivation. In experiment 1, glucose concentrations (0, 3.2, 6.25, 12.5, and 25%) were randomly presented during each test session. In experiment 2, individual glucose concentrations (0, 6.25, or 25%) were presented during separate sessions. For both, a deprivation condition was flanked by nondeprived (baseline) sessions. Concentration-response functions were comparable in both experiments. In each experiment, the shape of the concentration-response function was dramatically different during and after infusions. During infusions, there were no increases in glucose-elicited rhythmic oral responses beyond a very dilute concentration. After infusions, the concentration-response functions appeared linear across the concentration range. In both experiments, deprivation elevated responding only in the after-infusion periods. In experiment 1, the concentration-response function was uniformly elevated (on average, 27%) by deprivation, which if taken at face value would suggest an additive combination of taste and state feedback signals. In experiment 2, however, deprivation increased responding (approximately 30%) for 6.25%, but not for 0 or 25%, suggesting a stimulus specificity of the taste-state integration. Clearly then, the taste-state profiles differed as a function of experimental design. In the GENERAL DISCUSSION, we suggest that the uniform elevation of responding to all glucose concentrations, and to water, seen in experiment 1, may be an artifact of the random presentation of all stimuli during individual sessions. Experiment 2, in which stimuli were presented in a between-sessions design, may provide a truer reflection of the underlying integrative process.

Animals↗

Primate insular/opercular taste cortex: neuronal representations of the viscosity, fat texture, grittiness, temperature, and taste of foods.

It is shown that the primate primary taste cortex represents not only taste but also information about many nontaste properties of oral stimuli. Of 1,122 macaque anterior insular/frontal opercular neurons recorded, 62 (5.5%) responded to oral stimuli. Of the orally responsive neurons, some (53%) represented the viscosity, tested using carboxymethyl-cellulose in the range 1-10,000 cP. Other neurons (8%) responded to fat in the mouth by encoding its texture (as shown similar responses to nonfat oils), and 8% responded to gritty texture. Some neurons (35%) responded to the temperature of the liquid in the mouth. Some neurons responded to capsaicin, and others to fatty acids. Some neurons (56%) had taste responses. Some (50%) of these neurons were unimodal, responding to one of these types of stimulus, and the majority combined responsiveness to these types of stimulus, with 23% responding for example to both taste and temperature. Some neurons respond to taste, texture, and temperature unimodally, but others combine these inputs. None of these orally responsive neurons responded to odor or to the sight of food. These results provide fundamental evidence about the information channels used to represent the taste, texture, and temperature of food in the first cortical area involved in taste in the primate brain. The results are relevant to understanding the physiological and pathophysiological processes related to how the properties of oral stimuli are represented in the brain and thus to the control of food intake and food selection.

Action Potentials↗

Diet-induced plasticity in the taste system of an insect: localization to a single transduction pathway in an identified taste cell.

We studied exposure-induced sensitivity changes in an identified taste cell from Manduca sexta, a herbivorous caterpillar. This taste cell occurs within the lateral styloconic sensillum and responds selectively to compounds that humans characterize as bitter (e.g. caffeine, salicin and aristolochic acid). We made extracellular recordings from several classes of identified taste cell within the lateral sensillum, both before and after dietary exposure (for 48 h) to a suprathreshold concentration of caffeine, salicin or aristolochic acid. Our results revealed (1) that dietary exposure to caffeine desensitized the bitter-sensitive taste cell to caffeine, whereas dietary exposure to salicin or aristolochic acid did not desensitize the same taste cell to salicin or to aristolochic acid; (2) that dietary exposure to caffeine failed to alter the responsiveness of the sugar-, salt- or inositol-sensitive taste cells within the same sensillum; (3) that the caffeine-induced desensitization phenomenon generalized to salicin, a compound that stimulates the same transduction pathway as caffeine, but not to aristolochic acid, a compound that stimulates a different pathway; and (4) that chronically stimulating the lateral sensillum with caffeine, in the absence of ingestion, was sufficient to induce desensitization. We conclude that caffeine causes desensitization through a direct effect on a single transduction pathway within the bitter-sensitive taste cell.

Animals↗

[Studies on the taste perceptive threshold for 4 basic taste qualities at various sites of lingual surface].

Using filter-paper-disk method for 20 approximately 23 year-old 60 healthy females, taste perceptive thresholds for 4 basic taste qualities (sweetness, saltiness, sourness and bitterness) were measured detailedly at various sites of the lingual surface (lingual apex, lingual center, lingual margin, lingual radix). Sucrose, sodium chloride, sodium tartarate and quinine hydrochloride as taste-producing substances were used for the sweetness, saltiness, sourness and bitterness, respectively. The results revealed the apical site to be most sensitive to all of the sweetness, saltiness and sourness (and especially to the saltiness) and to be followed by marginal and radical sites in sensitiveness. The radical site was revealed to be most sensitive only to the bitterness, and the central site was remarkably hyposensitive to all of these taste qualities. These experimental results differ, in many respects, from those results of study which have been published by Kiesow in 1894 and are already established today (lingual sensitivity to taste qualities is found separately by site: sweetness, sourness and bitterness and highly perceptible at apical, marginal and radical sites, respectively, and saltiness is not very differently perceptible by site), indicating that further detailed reexamination will be needed to know how taste perceptive thresholds for 4 basic taste qualities are distributed in the lingual surface.

Adult↗

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↗

Taste responses to amino acids from facial nerve branches innervating oral and extra-oral taste buds in the channel catfish, Ictalurus punctatus.

Electrophysiological recordings were obtained from two branches of the facial nerve innervating oral and extra-oral taste receptive fields, respectively, in the channel catfish, Ictalurus punctatus. The results indicate that taste buds innervated by the ramus ophthalmicus superficialis (ros), innervating taste buds on the nasal barbel, and the ramus palatinus (rp), innervating taste buds on the anterior palate, have similar chemical specificities for amino acids. Among the amino acids tested, the most stimulatory compounds for both the ros and rp were L-alanine and L-arginine, having estimated electrophysiological thresholds of approximately 10(-9) M. Dose-response functions for amino acid stimuli recorded from both the ros and rp were power functions extending over 5 log units of stimulus concentrations. The general similarity in the nature of the taste input from spatially distinct gustatory areas supports a chemotactic role of the facial taste system in the channel catfish.

Adaptation, Physiological↗

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

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

Action Potentials↗