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Biochemical studies of taste sensation: II. Labelling of cyclic AMP of bovine taste papillae in response to sweet and bitter stimuli.

Labeling of cyclic AMP of tase papillae and its responsiveness to tast stimuli has been measured using whole papillae from bovine tongue prelabeled with [ 8-(14) C] adenine. Labeling was measured in circumvallate and fungiform papillae, both of which contain taste buds, and in filiform papillae and small blocks of tongue epithelium, which are devoid of taste buds. No differences were observed in the levels of activity. The labeling of cyclic AMP of circumvallate papillae showed only small increases (12-22%) in the presence of the taste stimulus sucrose (sweet), and the stimulatory effects were not statistically significant. The increase due to sucrose was not potentiated by theophylline. No stimulation by sucrose was observed with epithelium controls. Lactose, which is a poor taste stimulus, did not stimulate labeling of cyclic AMP in taste papillae. Theophylline, caffeine, and quinine (bitter) stimulated labeling of cyclic AMP by up to 2-fold, as did L-Epinephrine. Evidence for a specific role of cyclic AMP as a second messenger in taste sensation was not obtained. It is suggested that cyclic AMP might provide a mechanistic basis for studying some of the effects of sweet and bitter compounds in mixtures.

Adenine↗

Molecular structure influencing either a sweet or bitter taste among aldoximes.

A series of cyclohexylaldoximes was examined for their sweet or bitter taste using discriminant analysis. The structures of the molecules were described using molecular connectivity. A two-variable linear discriminant function and critical value were computed that correctly assigned 17 of the 20 molecules to their observed sweet or bitter taste categories. The same discriminant function can predict correctly the taste categories of seven of eight additional molecules.

Chemical Phenomena↗

Bitter substances suppress afferent responses to an appetitive mixture: evidence for peripheral integration of chemosensory stimuli.

The processes that lead from detection of chemicals, transduction, and coding with the appropriate message to initiate ingestion of a palatable meal or to reject a potentially noxious substance are poorly understood in vertebrates owing to the complex organization of the taste system. As a first step in elucidating the cellular basis of the behavioral differences elicited by appetitive stimuli and bitter compounds, we recorded from the afferent nerves conveying peripheral chemosensory information to the CNS in the head of the leech, Hirudo medicinalis. Superfusion of the chemosensory region of the lip of Hirudo with a mixture of NaCl (150 mM) and arginine (1 mM), an appetitive solution that elicits ingestion, increased the neuronal activity in the afferent cephalic nerves, for example (Zhang X, Wilson RJ, Li Y, Kleinhaus AL. 2000. Chemical and thermal stimuli have short-lived effects on the Retzius cell in the medicinal leech. J Neurobiol 43:304-311.). In the present paper we show that superfusing the lip with quinine or denatonium reduced the basal neural activity in the afferents. Furthermore, these bitter substances in the appetitive solution counteracted the increased activity the appetitive solution evoked in the cephalic nerves. Thus, the neural activity evoked by the application of appetitive and aversive stimuli to the chemosensory area of the lip paralleled the opposite behavioral responses to the same chemicals. The results suggest that individual leech taste cells possess receptors for both types of stimuli. Therefore, the leech may be a good model system in which to study peripheral taste events in cells that may possess multiple receptors and transduction mechanisms that interact to integrate information.

Action Potentials↗

Two antagonistic gustatory receptor neurons responding to sweet-salty and bitter taste in Drosophila.

In Drosophila, gustatory receptor neurons (GRNs) occur within hair-like structures called sensilla. Most taste sensilla house four GRNs, which have been named according to their preferred sensitivity to basic stimuli: water (W cell), sugars (S cell), salt at low concentration (L1 cell), and salt at high concentration (L2 cell). Labellar taste sensilla are classified into three types, l-, s-, and i-type, according to their length and location. Of these, l- and s-type labellar sensilla possess these four cells, but most i-type sensilla house only two GRNs. In i-type sensilla, we demonstrate here that the first GRN responds to sugar and to low concentrations of salt (10-50 mM NaCl). The second GRN detects a range of bitter compounds, among which strychnine is the most potent; and also to salt at high concentrations (over 400 mM NaCl). Neither type of GRN responds to water. The detection of feeding stimulants in i-type sensilla appears to be performed by one GRN with the combined properties of S+L1 cells, while the other GRN detects feeding inhibitors in a similar manner to bitter-sensitive L2 cells on the legs. These sensilla thus house two GRNs having an antagonistic effect on behavior, suggesting that the expression of taste receptors is segregated across them accordingly.

Action Potentials↗

Dietary effects of bitter gourd oil on blood and liver lipids of rats.

Bitter gourd is widely used as an edible plant in Asia. In this study, we evaluated the effects of bitter gourd oil (BGO) on the blood and liver lipids of rats. Three groups of rats were given a basal diet (AIN-93G) containing 7% fat by weight. The dietary fat consisted of soybean oil (control), soybean oil + BGO (6.5:0.5, w/w; 0.5% BGO), or soybean oil + BGO (5:2, w/w; 2.0% BGO). This fat treatment gave 3.4 and 15.4% of cis(c)9,trans(t)11,t13-18:3 in the dietary fat of 0.5 and 2.0% BGO, respectively. Fatty acid analysis showed the occurrence of c9,t11-18:2 in the liver of rats fed BGO diets, whereas this conjugated linoleic acid (CLA) isomer was not detected in the liver of rats fed the control diet. Furthermore, dietary BGO decreased the concentration of 18:2n-6 and increased the concentration of 22:6n-3. The formation of the CLA isomer in the liver lipids of rats fed BGO diets could be explained by either of the following two metabolic pathways, namely, enzymatic biohydrogenation of c9,t11,t13-18:3 or enzymatic isomerization of c9,c12-18:2. The BGO diets had significantly reduced free cholesterol levels with a trend toward an increase in HDL cholesterol, but there was no significant change in the total cholesterol. The dietary BGO also affected the level of plasma hydroperoxides. A slight but significant increase in hydroperoxides was found in the rats fed 2.0% BGO. This may be attributed to the lower oxidative stability of c9,t11,t13-18:3 in BGO.

Animal Nutritional Physiological Phenomena↗

Identification of coding single-nucleotide polymorphisms in human taste receptor genes involving bitter tasting.

T2Rs comprise a G-protein-coupled receptor superfamily that contains functionally defined bitter taste receptors. Here we report the tissue expressions and coding single-nucleotide polymorphisms (cSNPs) in human T2R genes (hT2R3, hT2R4, and hT2R5) on chromosome 7q31. We first demonstrated that hT2R3, hT2R4, and hT2R5 are actually expressed in the circumvallate papillae of the human tongue by reverse transcription-polymerase chain reaction (RT-PCR). We identified six cSNPs within the T2R receptor genes. The hT2R4 and hT2R5 contained four and one cSNPs that cause missense mutations, respectively, while hT2R3 included one silent nucleotide mutation. However, we could not find any nonsense mutations that resulted in a frameshift or a premature stop codon within the open reading frames. Genotype frequencies of each cSNP were in Hardy-Weinberg equilibrium. The identification of nucleotide diversity and amino acid polymorphisms in human T2R receptors could help clarify individual differences in the acceptability and sensitivity to bitter compounds.

Base Sequence↗

[Bitter peptides isolated from corn protein zein by hydrolysis with pepsin (author's transl)].

Eight peptides with bitter taste were isolated from a hydrolyzate of the corn protein zein using several chromatographic procedures. The peptides have the following amino acid sequences and taste thresholds (muM/ml): Ala-Ile-Ala (50-100), Ala-Ala-Leu (50-100), Gly-Ala-Leu (50-100), Leu-Glu-Leu (2.5-3.5), Leu-Glu-Leu (8-12), Leu-Val-Leu (1.5-2.5), Leu-Pro-Phe-Asn-Gln-Leu (0.1-0.2), Leu-Pro-Phe-Ser-Gln-Leu (0.1-0.2). The threshold for bitter taste decreases with increasing number of hydrophobic side chains (greater than or equal to C3) in the peptide. It increases in the presence of hydrophilic side chains to their polarity.

Amino Acid Sequence↗

[Contribution of linoleic acid to the bitter taste of poppy seeds (Papaver somniferum)].

In lipids isolated from poppy seeds which tasted "burning-bitter" the off-taste was associated with the free fatty acids fraction. In this fraction linoleic acid predominates, while oxidized fatty acids were among the minor constituents. The taste threshold of linoleic acid emulsified in water with monolinolein lies in the range of 4.0-6.0 mumol/ml. On the basis of its high concentration and relatively low taste threshold we conclude that free linoleic acid contributes significantly to the "burning-bitter" off-taste in poppy seeds.

Chromatography, Gel↗

Effect of bitter gourd and spent turmeric on constituents of glycosaminoglycans in different tissues in streptozotocin induced diabetic rats.

Diet is now one of the well established means in the management of diabetes. Bitter gourd and spent turmeric at 10% level were tested for their efficacy on glycosaminoglycan metabolism in various tissues viz., liver, spleen, lungs, heart and testis in control, diabetic and treated rats. The glycosaminoglycans (GAGs) were isolated from defatted and dried tissues. The contents of sulfated GAGs decreased in all the tissues and the decrease was more prominent in heart and testis. In the isolated GAGs, contents of total sugar, amino sugar, uronic acid and sulfate were studied. Decrease in total sugar content was maximum in testis. Amino sugar content decreased considerably in testis (38%) and lungs (15%). The content of uronic acid also decreased in testis (33%) besides heart (29%) and liver (25%). Sulfate groups in GAGs perform pivotal functions in many biological events and decrease in sulfate content was significant in heart (40%), testis (37%) and liver (37%). GAGs profile on the cellulose acetate electrophoresis revealed that heparan sulfate (HS), hyaluronic acid (HA) and chondroitin sulfate/dermatan sulfate (CS/DS) were present in liver, spleen and lungs. HS, CS were present in heart, DS/CS was observed in testis. The observed beneficial effects in GAGs metabolism during diabetes may be due to the presence of high amounts of dietary fibres present in bitter gourd and spent turmeric, besides, possible presence of bioactive compounds in one or both of them.

Amino Sugars↗

Receptor potential of the frog taste cell in response to bitter stimuli.

Molecular mechanisms of generation of the receptor potential in the bullfrog taste cell for bitter stimuli were investigated with an intracellular recording technique. During generation of the receptor potential in response to bitter stimuli, the input resistance of the taste cell increased slightly. We could not find the reversal potential for the depolarizing receptor potential induced by quinine-HCl(Q-HCl). The Q-HCl-induced response was increased with decreasing Cl- concentration in the superficial fluid. The Q-HCl response was greatly reduced by interstitial furosemide, as a blocker of Na+/Cl- cotransport, indicating that a Na+/Cl- cotransport occurs through the basolateral membrane of Q-HCl sensitive taste cells. Therefore it is concluded that the receptor potential for Q-HCl stimulation is produced by an active secretion of intracellularly accumulated Cl- through Cl- pumps of the apical receptive membrane.

Animals↗

Denatonium bitter tasting among transgenic mice expressing rat von Ebner's gland protein.

Von Ebner's gland protein (VEGP) is a secretory protein, which is abundantly expressed in the small von Ebner's salivary glands of the tongue. VEGP as component of the perireceptor environment around taste papillae might function as transporter of hydrophobic molecules, for example bitter substances. Here we report a new approach to investigate the physiological role of VEGP by expression of the cloned rat VEGP gene in transgenic mice. Taste papillae of mice, in contrast to rats, do not contain VEGP. The founder mouse 4345 and three offspring carry the transgene as shown by PCR analysis and saliva of the transgenic mice contains high amounts of VEGP. In two-bottle preference tests, transgenic and nontransgenic siblings show significantly different capabilities to taste the bitter compound denatonium benzoate at 10 microM. The reduced sensitivity of transgenic mice to denatonium benzoate points to a clearance function of VEGP the specificity of which for taste compounds and other molecules remains to be seen.

Animals↗

The cytotoxic and cytostatic effects of the bitter melon (Momordica charantia) on human lymphocytes.

We have previously reported that a crude extract from the bitter melon (Momordica charantia) killed human leukemic lymphocytes in a dose-dependent manner while not affecting the viability of normal human lymphocyte cells at these same doses (Takemoto et al., 1980). We now report that the crude preparation has both cytostatic and cytotoxic activities which are heat stable and trypsin-sensitive. Time and dose-response curves suggest that the factors act quickly, perhaps by entry into the cell. The effects of the crude extract are complete after only 2 hr of exposure. These activities are not due to the presence of the lectins from bitter melon seeds, as these purified proteins had no activity against human lymphocytic cells.

Animals↗

Microbial degradation of amygdalin of bitter apricot seeds (Prunus armeniaca).

Amygdalin is a cyanogenic glycoside occurring among others in almonds and bitter apricot seeds with interesting levels of dietary protein. Utilization of seeds for human or animal nutrition requires adequate detoxification. In the present paper, selected filamentous fungi (Mucor circinelloides, Penicillium nalgiovense) and yeasts (Hanseniaspora valbyensis, Endomyces fibuliger) were tested for their in-situ ability to decompose amygdalin. The latter (Endomyces fibuliger) was best able to grow on autoclaved bitter apricot seeds and detoxify them from 30 microMol CN/g dry matter to less than 1 microMol CN/g dry matter after 48 h of incubation at 27 degrees C.

Amygdalin↗

Consumption of alcohol compared to another bitter solution in a limited access drinking paradigm.

A number of recent studies have shown that free-feeding rats will consume pharmacologically significant amounts of alcohol when given access to alcohol for short periods of time daily. The present experiment was designed to examine the role of taste in inducing alcohol consumption in this type of periodic availability paradigm, by comparing amounts drunk thereby to amounts of another bitter solution, sucrose octa acetate (SOA), presented on a similar schedule. In contrast to alcohol, which was consistently preferred to water, at a concentration as high as 12% (w/v), preference for SOA diminished across days, indicating that the bitter taste alone was insufficient to increase consumption of this solution. The results also suggest the animals may be drinking in response to external conditioning stimuli, an effect which has previously been demonstrated in regard to feeding.

Alcohol Drinking↗

Potential of immobilized bitter gourd (Momordica charantia) peroxidases in the decolorization and removal of textile dyes from polluted wastewater and dyeing effluent.

Immobilized peroxidases from Momordica charantia were highly effective in decolorizing reactive textile dyes compared to its soluble counterpart. Dye solutions, 50-200 mg/l, were treated with soluble and immobilized bitter gourd peroxidases (specific activity of 99.0 EU per mg protein). The decolorization of dyes with soluble and immobilized enzyme was maximum in the range of pH 3.0-4.0. The effect of different temperatures on the dye decolorization was monitored and it was observed that all the dyes were maximally decolorized at 40 degrees C. In order to examine the operational stability of the immobilized preparation, the enzyme was repeatedly exploited for the decolorization of the dyes from fresh batch of dye solutions. Even after 10 cycles in each case the immobilized preparation retained nearly 50% of the initial enzyme activity. The immobilized enzyme exhibited more than 90% of the original activity while the soluble enzyme lost 33% of the initial activity when stored for 40 d at room temperature. Mixtures of three, four and eight dyes were prepared and treated with soluble and immobilized bitter gourd peroxidase. Each mixture was decolorized by more than 80% when treated with immobilized enzyme. Dyeing effluent collected from local dyers was treated with both types of enzyme preparations. Immobilized enzyme was capable of removing remarkably high concentration of color from the effluent. TOC content of soluble and immobilized enzyme treated individual dyes, mixture of dyes and dyeing effluent was determined and it was observed that higher TOC was removed after treatment with immobilized enzyme.

Buffers↗

Receptors for bitter, sweet and umami taste couple to inhibitory G protein signaling pathways.

Taste receptors are thought to couple to the G protein Galpha-gustducin to initiate signal transduction cascades leading to taste perception. To further characterize the G protein-coupling selectivity of these receptors, we expressed them in HEK293 cells and monitored the modulation of different signaling pathways upon stimulation. We found that the bitter compound cycloheximide induces phosphorylation of extracellular signal-regulated kinases1 and 2 (ERK 1/2) and inhibits cAMP accumulation in HEK293 cells expressing the mouse bitter T2R(5) receptor. These effects are totally abolished upon treatment with pertussis toxin. On the other hand, sweeteners and monosodium glutamate induce phosphorylation of ERK1/2 and inhibit cAMP accumulation in HEK293 cells expressing the human sweet T1R(2)/T1R(3) receptor and the human umami T1R(1)/T1R(3) receptor, respectively. The effects of these taste modalities are also prevented by treatment with pertussis toxin. Collectively, our results show that taste receptors can functionally couple to Galpha(i/o) proteins to transmit intracellular signals.

Adenylyl Cyclase Inhibitors↗

Effects of sweet and bitter gustatory stimuli in anorexia nervosa on EEG frequency spectra.

The possible differences in processing gustatory stimuli in anorexic patients compared to healthy control subjects was investigated by electrophysiological methods. The electroencephalogram (EEG) was recorded in outpatients treated with anorexia nervosa (AN) and age-matched controls after exposure to sweet (milk chocolate) and bitter (black tea) taste stimuli. Power spectrum analysis was performed on EEG epochs recorded in the above conditions. Compared to controls a significantly higher percent of theta, and lower percent of alpha1 band power was found in anorexic patients, irrespective of the kind of taste effects and hemispheric side. The pattern of activation caused by sweet and bitter stimuli was found to be different in these two groups, possibly indicating altered gustatory processing mechanisms in AN.

Adolescent↗

Diverse tastes: Genetics of sweet and bitter perception.

Humans will eat almost anything, from caribou livers to rutabagas, but there are some types of foods, and their associated taste qualities, that are preferred by large groups of people regardless of culture or experience. When many choices are available, humans chose foods that taste good, that is, create pleasing sensations in the mouth. The concept of good taste for most people encompasses both flavor and texture of food, and these sensations merge with taste proper to form the concept of goodness. Although we acknowledge the universality of the goodness (sweet) or badness (bitter) of basic taste qualities, we also find that people differ, sometimes extremely so, in their ability to perceive and enjoy these qualities and, by extension, food and drink. The reasons for these differences among people are not clear but are probably due to a combination of experience beginning at an early age, perhaps in utero; learning, for example, as with conditioned taste aversions; sex and maturity; and perceptual differences that arise from genetic variation. In this review, we focus on individual variations that arise from genetic differences and review two domains of science: recent developments in the molecular biology of taste transduction, with a focus on the genes involved and second, studies that examine biological relatives to determine the heritability of taste perception. Because the receptors for sweet, savory (umami), and bitter have recently been discovered, we summarize what is known about their function by reviewing the effect of naturally occurring and man-made alleles of these receptors, their shape and function based on receptor modeling techniques, and how they differ across animal species that vary in their ability to taste certain qualities. We discuss this literature in the context of how taste genes may differ among people and give rise to individuated taste experience, and what is currently known about the genetic effects on taste perception in humans.

Animals↗