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Biochemical studies of taste sensation. XIII. Enantiomeric specificity of alanine taste receptor sites in catfish, Ictalurus punctatus.

Specific binding of amino acid taste stimuli is known to occur to a sedimentable fraction (P2) from catfish (Ictalurus punctatus) taste epithelium or to purified plasma membranes from that fraction. L-Alanine, a potent taste stimulus for the catfish, binds in a reversible and saturable manner to these preparations. The extent to which the enantiomeric stimuli, L- and D-alanine, interact with the same or different receptor/transduction processes is investigated here both electrophysiologically and biochemically. With an electrophysiological assay, L-alanine was the more potent stimulus across a concentration range of 10(-9)-10(-3) M, yet both enantiomers displayed approximately the same threshold. The concentration-electrophysiological response functions for each enantiomer were different. That of L-alanine was approximately linear across the (log) concentration range while that of D-alanine was non-linear, with small but definitely observable responses being noted from 10(-9)-10(-5) M D-alanine, and larger incremental responses thereafter. With most of the nerve bundle preparations studies, L- and D-alanine cross-adapted one another, but this cross-adaptation was not always complete. Experiments in which both L- and D-alanine were present in a 1:1 mixture of equally stimulatory concentrations suggested the existence of receptor or transduction processes unique to each enantiomer. Biochemically binding studies demonstrated high affinity binding sites for both enantiomers with values of Kd-app for L-alanine of 1.5 microM and for D-alanine of 25 microM. For both enantiomers, additional lower-affinity binding sites were observable. The capacity of the lower-affinity sites was particularly great for D-alanine. The enantiomers competed one with the other for binding, with L-alanine showing greater competitive ability than D-alanine at low concentrations. For the high affinity sites, double-reciprocal plots of the data suggested a competitive mechanism. The lower affinity sites for D-alanine were less accessible to L-alanine compared with the high affinity sites of D-alanine. Both the biochemical and electrophysiological results indicate that while a portion of the responses to L- and D-alanine occurs through a common receptor/transduction process, there exist independent receptor/transduction processes for the enantiomers, L- and D-alanine.

Alanine↗

Astringency of bovine milk whey protein.

Whey protein solutions at pH 3.5 elicited an astringent taste sensation. The astringency of whey protein isolate (WPI), the process whey protein (PWP) that was prepared by heating WPI at pH 7.0, and the process whey protein prepared at pH 3.5 (aPWP) were adjusted to pH 3.5 and evaluated by 2 sensory analyses (the threshold method and the scalar scoring method) and an instrumental analysis (taste sensor method). The taste-stimulating effects of bovine and porcine gelatin were also evaluated. The threshold value of astringency of WPI, PWP, and aPWP was 1.5, 1.0, and 0.7 mg/mL, respectively, whereas the gelatins did not give definite astringency. It was confirmed by the scalar scoring method that the astringency of these proteins increased with the increase in protein concentration, and these proteins elicited strong astringency at 10 mg/mL under acidic conditions. On the other hand, the astringency was not elicited at pH 3.5 by 2 types of gelatin. A taste sensor gave specific values for whey proteins at pH 3.5, which corresponded well to those obtained by the sensory analysis. Elicitation of astringency induced by whey protein under acidic conditions would be caused by aggregation and precipitation of protein molecules in the mouth.

Animals↗

Gastrointestinal reactivity in rats lacking anterior insular neocortex.

Behavioral and physiological studies have shown that the insular neocortex participates in a wide variety of special visceral processes, in particular, the higher-order integration of taste stimuli and the regulation of autonomic activity. The present experiment examined the involvement of the anterior insular (gustatory) neocortex (AIGN) in gastrointestinal reactivity and taste learning in rats by the use of a modified taste aversion training procedure. Normal rats and rats lacking AIGN demonstrated similar "illness thresholds" to the early onset symptoms of ingested LiCl. Conversely, animals lacking AIGN showed significant impairments in taste aversion learning ability. From a consideration of several research findings it is concluded that animals lacking AIGN can normally perceive tastes and illness, but these animals experience reliable impairments in taste aversion learning ability.

Animals↗

[A study of changes in gustatory sense during pregnancy].

A change in gustatory function is generally well known as one of the manifestations in the early stage of pregnancy. However, there have been comparatively few studies of the changes in taste and gustatory function resulting from pregnancy. In particular, so far there have been no reports of investigations of two or more pregnant women in which gustatory sense in the same pregnant woman was examined serially over time. In this study, gustatory tests were performed over time on pregnant women having a normal course, so as to examine in detail the changes in gustatory function resulting from pregnancy. Selected for this study were 97 healthy pregnant women (ranging in age from 21 to 36 years, with an average of 27.9 +/- 3.2 years). Gustatory tests were performed by electrogustometry and the filter-paper disk method over the areas of the chorda tympani nerve and glossopharyngeal nerve. Of the 97 subjects, gustatory tests could be performed serially with time throughout the duration of pregnancy in 32 subjects (from 21 to 34 years old, with an average age of 28.3 +/- 2.9 years), and changes in gustatory sense with progress in pregnancy were analyzed statistically. In addition, serum trace elements were measured in 72 of the subjects. Thirty non-pregnant healthy women (from 18 to 34 years old, with an average age of 24.8 +/- 4.3 years) were used as control. The gustatory test was performed by the same person in all cases. An increase in gustatory threshold was observed in pregnant women as compared with that of non-pregnant women. Especially, a tendency of a marked decrease was noted in gustatory function in the first trimester. A marked change in secretion of hormones in the early stage of pregnancy is considered to be the underlying cause of such a decrease in gustatory function. The serum zinc level was in the normal range in the early stage of pregnancy, but it tended to decrease over the period from the midtrimester to the third trimester. Therefore, it is difficult to explain dysgeusia in the early stage of pregnancy as being associated with a deficiency of zinc.

Adult↗

Detection of tastes in mixture with other tastes: issues of masking and aging.

When one taste (masker) is strong enough, it can completely mask another task (target) of different quality. How strong the masker must be to do this depends on how strong the target is. As the target concentration is increased, the masking concentration must be increased, too, but in ever-increasing proportion. To quantify the conditions for such complete masking, the target's detection threshold was measured as a function of the masker's concentration, from zero to strong. This was done for 12 binary combinations of sucrose, sodium chloride, citric acid and quinine hydrochloride. The 12 functions generated show that some tastants mask each other much more efficiently than others. Masking gives new insight into the role of aging in taste: older (66-90 years) subjects' thresholds, regardless of masking concentration, always measured a constant factor higher than younger (18-29 years) subjects' thresholds (about two to seven times higher, depending on target tastant). Thus, with increasing level of the masker, the thresholds of young and elderly go up in parallel. Thresholds of tastants in water alone are false predictors of elderly persons' ability to perceive ingredients like salt and sugar condiments in foods, where, because of masking, their thresholds can be several times higher than in water. Age manifested itself relatively mildly in sucrose and citric acid, moderately in sodium chloride, and strongly in quinine hydrochloride.

Adolescent↗

Taste receptor mechanism of salts in frog and rat.

Salts carrying impermeable organic cations elicited large responses in the frog and rat taste nerves. Amiloride suppressed the responses of the frog and rat taste nerves to NaCl, KCl and the organic salts to a similar extent. The estimated equilibrium potentials for Na+ and K+ at the thresholds determined were more negative than the resting potentials of the taste cells. The results obtained suggest that the taste responses to salts including NaCl are not induced by entry of cations via apical membranes of taste cells.

Amiloride↗

Smell and taste disorders.

Disorders of taste and smell can present a challenge to the facial plastic surgeon. Obtaining a detailed history and examination is the key to the diagnosis and work-up of olfactory and gustatory dysfunction. Easy-to-administer tests are available for olfactory evaluation(University of Pennsylvania Smell Identification Test) and gustatory (taste sticks, tasting tablets) evaluation. The prognosis and management of olfactory and gustatory disease depend on its etiology. Despite ongoing research, the treatment of the disorders of smell and taste is limited.

Chemoreceptor Cells↗

Amiloride sensitivity of the chorda tympani response to sodium chloride in sodium-depleted Wistar rats.

Peripheral gustatory mechanisms that may contribute to the expression of sodium (Na) appetite have been a focus of interest for many years. Because amiloride-sensitive Na transport is involved in the generation of neural signals in response to NaCl stimulation, the present study assessed whether changes in amiloride sensitivity of the neural response to NaCl accompany the induction of a Na appetite in the rat. Na deprivation was achieved by acute depletion with the diuretic furosemide. The magnitude of the whole-nerve chorda tympani response to 0.5 M NaCl was reduced in Na-depleted, compared with Na-replete, rats, which provides qualified support for previous reports that the induction of a Na appetite is associated with reduced neural responses to NaCl. However, changes in sensitivity to the specific Na channel blocker amiloride hydrochloride as a result of Na depletion were not evident. These findings suggest that the behavioral and neural changes that occur after Na depletion are not based on changes in amiloride sensitivity in the taste bud.

Amiloride↗

The mammalian amiloride-insensitive non-specific salt taste receptor is a vanilloid receptor-1 variant.

The amiloride-insensitive salt taste receptor is the predominant transducer of salt taste in some mammalian species, including humans. The physiological, pharmacological and biochemical properties of the amiloride-insensitive salt taste receptor were investigated by RT-PCR, by the measurement of unilateral apical Na+ fluxes in polarized rat fungiform taste receptor cells and by chorda tympani taste nerve recordings. The chorda tympani responses to NaCl, KCl, NH4Cl and CaCl2 were recorded in Sprague-Dawley rats, and in wild-type and vanilloid receptor-1 (VR-1) knockout mice. The chorda tympani responses to mineral salts were monitored in the presence of vanilloids (resiniferatoxin and capsaicin), VR-1 antagonists (capsazepine and SB-366791), and at elevated temperatures. The results indicate that the amiloride-insensitive salt taste receptor is a constitutively active non-selective cation channel derived from the VR-1 gene. It accounts for all of the amiloride-insensitive chorda tympani taste nerve response to Na+ salts and part of the response to K+, NH4+ and Ca2+ salts. It is activated by vanilloids and temperature (> 38 degrees C), and is inhibited by VR-1 antagonists. In the presence of vanilloids, external pH and ATP lower the temperature threshold of the channel. This allows for increased salt taste sensitivity without an increase in temperature. VR-1 knockout mice demonstrate no functional amiloride-insensitive salt taste receptor and no salt taste sensitivity to vanilloids and temperature. We conclude that the mammalian non-specific salt taste receptor is a VR-1 variant.

Amiloride↗

Enhanced gustatory neural responses to sugars in the diabetic db/db mouse.

Sweet taste sensitivity in a genetic model of diabetes, the db/db mouse, in which a single major gene defect leads to the expression of diabetes and obesity, was studied by examining chorda tympani nerve responses to various taste stimuli, including sugars. The chorda tympani responses to four sugars, sucrose, fructose, glucose, and maltose, in adult db/db mice showed greater relative magnitudes and lower thresholds than those in adult lean mice, but responses to other basic taste stimuli, such as NaCl, HCl, and quinine HCl, were not different in the two groups. Behavioral experiments using a two-bottle preference test demonstrated that taste preference scores for the four sugars at suprathreshold concentrations, except 1.0 M, were higher in db/db than in control mice. Infant mice of 7-9 days of age possessing the genotype db/db also exhibited greater neural responses and lower thresholds for sugars than infant control mice, whereas streptozotocin-induced adult diabetic mice possessing the genotype +/+ did not exhibit larger sugar responses. These findings suggest that the enhanced sugar sensitivities observed in db/db mice are probably determined by a single major gene, db. The db gene may act on a common factor(s) involved in the stimulus-secretion coupling in the pancreatic B cell and the taste cell of db/db mice.

Aging↗

Hedonic and sensory characteristics of odors conditioned by pairing with tastants in humans.

Animals readily acquire positive odor-taste hedonic associations, but evidence for this in humans remains weak and was explored further. Retronasal pairing of odors with sucrose or salty stimuli (Experiment 1) increased the rated sweetness of sucrose-paired odors without altering liking, although changes in odor pleasantness correlated with sucrose liking. Experience of odors with sucrose or quinine by sweet likers (Experiment 2) found increased pleasantness and sweetness for sucrose-paired odors, whereas quinine-paired odors became less liked and more bitter. Odor-sucrose pairings in sweet likers and dislikers (Experiment 3) found increased sweetness in both groups but increased odor liking only in likers. These data suggest that evaluative and sensory learning are dissociable and that evaluative changes are sensitive to individual differences in sweet liking.

Adult↗

Whole nerve chorda tympani responses to sweeteners in C57BL/6ByJ and 129P3/J mice.

The C57BL/6ByJ (B6) strain of mice exhibits higher preferences than does the 129P3/J (129) strain for a variety of sweet tasting compounds. We measured gustatory afferent responses of the whole chorda tympani nerve in these two strains using a broad array of sweeteners and other taste stimuli. Neural responses were greater in B6 than in 129 mice to the sugars sucrose and maltose, the polyol D-sorbitol and the non-caloric sweeteners Na saccharin, acesulfame-K, SC-45647 and sucralose. Lower neural response thresholds were also observed in the B6 strain for most of these stimuli. The strains did not differ in their neural responses to amino acids that are thought to taste sweet to mice, with the exception of L-proline, which evoked larger responses in the B6 strain. Aspartame and thaumatin, which taste sweet to humans but are not strongly preferred by B6 or 129 mice, did not evoke neural responses that exceeded threshold in either strain. The strains generally did not differ in their neural responses to NaCl, quinine and HCl. Thus, variation between the B6 and 129 strains in the peripheral gustatory system may contribute to differences in their consumption of many sweeteners.

Animals↗

Mouse taste cells with G protein-coupled taste receptors lack voltage-gated calcium channels and SNAP-25.

BACKGROUND: Taste receptor cells are responsible for transducing chemical stimuli from the environment and relaying information to the nervous system. Bitter, sweet and umami stimuli utilize G-protein coupled receptors which activate the phospholipase C (PLC) signaling pathway in Type II taste cells. However, it is not known how these cells communicate with the nervous system. Previous studies have shown that the subset of taste cells that expresses the T2R bitter receptors lack voltage-gated Ca2+ channels, which are normally required for synaptic transmission at conventional synapses. Here we use two lines of transgenic mice expressing green fluorescent protein (GFP) from two taste-specific promoters to examine Ca2+ signaling in subsets of Type II cells: T1R3-GFP mice were used to identify sweet- and umami-sensitive taste cells, while TRPM5-GFP mice were used to identify all cells that utilize the PLC signaling pathway for transduction. Voltage-gated Ca2+ currents were assessed with Ca2+ imaging and whole cell recording, while immunocytochemistry was used to detect expression of SNAP-25, a presynaptic SNARE protein that is associated with conventional synapses in taste cells. RESULTS: Depolarization with high K+ resulted in an increase in intracellular Ca2+ in a small subset of non-GFP labeled cells of both transgenic mouse lines. In contrast, no depolarization-evoked Ca2+ responses were observed in GFP-expressing taste cells of either genotype, but GFP-labeled cells responded to the PLC activator m-3M3FBS, suggesting that these cells were viable. Whole cell recording indicated that the GFP-labeled cells of both genotypes had small voltage-dependent Na+ and K+ currents, but no evidence of Ca2+ currents. A subset of non-GFP labeled taste cells exhibited large voltage-dependent Na+ and K+ currents and a high threshold voltage-gated Ca2+ current. Immunocytochemistry indicated that SNAP-25 was expressed in a separate population of taste cells from those expressing T1R3 or TRPM5. These data indicate that G protein-coupled taste receptors and conventional synaptic signaling mechanisms are expressed in separate populations of taste cells. CONCLUSION: The taste receptor cells responsible for the transduction of bitter, sweet, and umami stimuli are unlikely to communicate with nerve fibers by using conventional chemical synapses.

Animals↗

Clinical assessment of retronasal olfactory function.

OBJECTIVES: To develop a test kit for the simple assessment of retronasal olfactory function and to compare orthonasal and retronasal olfactory function in healthy subjects and patients with olfactory disorders. DESIGN AND PATIENTS: We tested 230 individuals with normosmia, hyposmia, and anosmia using grocery-available powders. Initially, 30 different substances were investigated. Subjects identified each substance using a list with 4 verbal items (forced choice). After preliminary experiments, 20 items were selected according to the degree to which they were identified by normosmic and anosmic subjects. Orthonasal olfactory function was assessed psychophysically using "sniffin' sticks," which includes tests for odor identification, discrimination, and butanol odor thresholds. In addition, anosmia was confirmed electrophysiologically by means of olfactory-evoked potentials. RESULTS: In healthy subjects, there was a test-retest reliability correlation of r(27) = 0.76 for retronasal olfactory function, which is similar to other odor identification tests. Retronasal testing in normosmic subjects allowed for the discrimination of sex-related differences, with women scoring higher than men (P =.007), and the identification of a slight decrease with age (r(120) = -0.20; P =.03). Orthonasal and retronasal identification of odors was found to correlate (r(86) = 0.78; P<.001). Retronasal testing allowed for the discrimination between normosmia, hyposmia, and anosmia (P<.001). In addition, retronasal performance of anosmic patients appeared to improve with duration of anosmia (P =.03). No difference was found between patients with anosmia of different origin. CONCLUSION: Results of the present investigation indicate that the assessment of retronasal olfactory function is possible using oral stimulus presentation.

Adolescent↗

Discriminative feature integration by individuals.

A tactile pattern on the skin generally comes to be recognised from its function within action. What is perceived tactually and may generate a textural or geometrical sensation is therefore a set of dynamic characteristics that can be extracted by mechanoreceptors and central neural processing from the physics of the interactions of hands, mouth or other parts of the body with common objects and materials. The recognised dynamic (as is true of any sensory modality) may be diagnosable as distinct features that are integrated by the whole system's operation. If so, this transmission of feature information over a channel or in a dimension can be measured as the strength of the influence of that aspect of the stimulating pattern on the observed output, in units of the acuity of response differences for deviations of the pattern from its familiar configuration of features. When applied at the individual level, such multidimensional discrimination analysis provides powerful diagnoses of pattern-recognition processes.

Attention↗