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Biochemical studies of taste sensation. VII. Enhancement of taste stimulus binding to a catfish taste receptor preparation by prior exposure to the stimulus.

The taste receptor membrane fraction (Fraction P2) was prepared from a homogenate of the taste tissue of the channel catfish Ictalurus punctatus. This included the rostral, dorsal, and dorsolateral surfaces of the catfish in addition to those of the barbels. The yield of Fraction P2 is 4-7 mg protein from an individual fish, with a purification averaging 8- to 15-fold over that of the crude whole homogenate and essentially quantitative recovery of binding activity in Fraction P2. Treatment of Fraction P2 in vitro with a high concentration of the taste stimulus molecule L-alanine led to a several-fold enhancement of binding activity. Enhancement of the binding of 3H L-alanine was observed after treatment with unlabeled 10 mM L-alanine and removal of the L-alanine by washing. Enhancement occurred whether the preparation was stored frozen (-65 degrees C) for an extended period in the presence of the L-alanine, or merely exposed to it in the cold without freezing. D-Alanine enhanced the binding activity of 3H L-alanine to about 60% of the level induced by L-alanine. Nonspecific binding of 3H L-alanine was unaffected by the treatment. Scatchard analyses of saturation curves for binding of 3H L-alanine to freshly prepared Fraction P2 and to L-alanine-treated Fraction P2 revealed no change in the KD value, but a several-fold increase occurred in the amount bound. Binding activity is operationally defined. Because the enhancement observed here is reminiscent of an increase in transport due to a countertransport effect, further studies were carried out to examine whether the phenomenon reflects transport or true binding. The measured binding was not increased in the presence of Na+, indicating that it is not due to an Na+-coupled transport of L-alanine. When Fraction P2 was preloaded with L-alanine (10(-6)--10(-2) M) prior to assay, no stimulation of binding was observed; instead, binding decreased. This result is consistent with a true binding phenomenon but not with a carrier-mediated transport process to explain the enhancement phenomenon. Binding assays carried out over a range of osmolarities revealed decreased binding at high osmotic strengths, suggesting that a significant portion of the ligand might be contained in vesicles. It is postulated that "hidden" or "buried" receptor sites exist in the Fraction P2 as isolated, and that these are exposed upon perturbation of the membrane structure by a high ligand concentration.

Alanine

Biochemical studies of taste sensation--VIII. Partial characterization of alanine-binding taste receptor sites of catfish Ictalurus punctatus using mercurials, sulfhydryl reagents, trypsin and phospholipase C.

1. Taste receptors for L-alanine in the channel catfish Ictalurus punctatus have been partially characterized. The binding activity, which is localized to a sedimentable fraction (Fraction P2), was assayed with L-[3H]alanine as the ligand. 2. Addition of HgCl2 or p-mercuribenzoate to the assay at 0.1-1 mM markedly inhibited binding. The effect was not reversible and was unaffected by increased L-alanine in the binding assay. 3. The sulfhydryl reagents iodoacetate, 5,5'-dithiobis(2-nitrobenzoic acid), arsenite, and N-ethylmaleimide did not show appreciable inhibition of binding. The results suggest that the inhibitory effect of mercurials is not on specific sulfhydryl groups at alanine-binding sites. 4. Treatment of Fraction P2 with phospholipase C decreased binding activity and treatment with trypsin led to increased binding activity.

Alanine

[Analysis of the effect of bicarbonate on taste receptor sensitivity to organic acids].

Addition of NaHCO3 or NaOH to the solutions of lactic or pyruvic acids decreased threshold concentrations of H+ ions and increased intensity of afferent impulsation in the cat chroda tympani nerve. In spite of the fact that the concentrations of both lactate and pyruvate anions with the presence of NaOH and NaHCO3 are the same at equal pH values the observed changes of the receptors responses were more intensive if NaHCO3 had been added. Therefore the influence of NaHCO3 on the perception of organic acids depends not only on its buffer properties but, probably, on direct action of CO2 appearing as the result of interaction of NaHCO3 and acids, as well. This assumption was confirmed by the data on high sensitivity of the cat tongue receptors for the carbon dioxide.

Animals

Three-dimensional mapping of the sweet taste receptor site.

The active sites of the receptors for sweet and bitter tastants are shown to be related by a simple symmetry operation. This relationship, in turn, allows the identification of the critical geometrical features of both receptor sites. The model proposed for the sweet site is shown to be consistent with a large number of (conformationally rigid) sweet molecules.

Models, Molecular

[Relationship between the anionic structure of organic acids and the response of cat taste receptors].

With the aid of analysis of afferent impulse activity in the cat chorda tympani, it was shown that the effect of application of organic acids solutions of the same pH to the tongue could be represented as follows: propionic acid greater than lactic acid greater than pyruvic acid. These data confirm the suggestion that carboxylic acids without polar groups are the most effective, and an anion which has OH-group in alpha-position is more effective than an anion with keto-group.

Animals

[Differences in the responses of taste receptors to organic and inorganic acids with changes in the concentration of bicarbonate in the solution].

The thresholds of the pH for citric acid (pH=4.9) were found to exceed by 1.4 pH the thresholds for HC1 (3.5) at 1.2 mmol/1 bicarbonate in the solution. The reaction to citric acid was higher than to HC1 at equal pH. Decreasing of bicarbonate from 1.2 mmol/1 to 0 reduced pH threshold only for sitric acid from 4.90 to 3.15. pH threshold for HC1 remained 3.5 The chorda tympani response to stimulation with solutions containing bicarbonate (1.2 mmol/1) was higher than in absence of bicarbonate. The data obtained suggest two ranges of the acids in action.

Animals

Dietary soybean or seaweed (Kappaphycus sp.) modulates taste-related gene (tas1r1 and tas1r2.2) expression in Nile tilapia (Oreochromis niloticus).

Taste perception plays a central role in fish feeding behaviour by influencing feed recognition, intake, and nutrient sensing. As aquaculture increasingly adopts plant-based ingredients to replace fishmeal, understanding how these diets affect gustatory mechanisms is critical. This study evaluated TAS-family taste receptor genes in Nile tilapia (Oreochromis niloticus) fed nutritionally formulated diets containing fishmeal (TFM), soybean meal (TSB), red seaweed (Kappaphycus sp., TSW), alongside a natural meal (TNM) reference. Fish were reared for 62 days under controlled conditions, after which growth performance was evaluated, and tongue transcriptomes were analysed using RNA sequencing to identify diet-associated gene expression changes. Candidate TAS1R and TAS2R receptors were identified through conserved domain screening and phylogenetic validation, and differential expression analysis was performed using DESeq2. Growth performance did not differ significantly among diets, although the soybean group showed the highest weight gain. Under the conditions of this study, most taste-related genes remained transcriptionally stable across formulated diets, indicating limited responsiveness of the gustatory system to ingredient substitution. Transcriptional differences were mainly observed in comparisons involving the TNM (TSB vs TNM and TSW vs TNM), where tas1r1 was upregulated, suggesting altered amino acid sensing relative to the non-formulated diet. Among formulated diets, tas1r2.2 was upregulated in the TSW vs TFM comparison, indicating potential modulation of carbohydrate-related taste pathways associated with seaweed inclusion. No reliable TAS2R transcripts were detected, likely due to low expression or tissue-specific distribution. Overall, taste receptor expression in Nile tilapia appears resilient to dietary variation, with selective modulation of TAS1R genes providing molecular insight into chemosensory adaptation to sustainable feed ingredients.

Animals

Biochemical studies of taste sensation: binding to taste tissue of 3H-labeled monellin, a sweet-tasting protein.

Binding of 3H-labeled methylated monellin to taste receptor tissue was demonstrated in vitro. Preparation of bovine and human circumvallate (taste) papillae bound more of the ligand than did lingual and nonlingual epithelial preparations devoid of taste buds. Binding to the taste preparations saturated at high ligand concentrations. Furthermore, sugars and other sweet-tasting molecules appeared to compete to some extent with this sweet-tasting protein for its binding sites. These binding measurements of the intensity sweet-tasting protein monellin to taste receptor preparations help to establish the binding interactions as an initial step taste sensation.

Animals