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PubMed2001

Molecular genetic identification of a candidate receptor gene for sweet taste.

A cDNA clone encoding a novel member of the putative taste receptor T1R family, designated T1R3, was isolated from circumvallate papillae of the mouse tongue using degenerate primers. Reverse transcription-polymerase chain reaction analysis showed predominant expression of the receptor in circumvallate papillae. In situ hybridization analysis revealed that T1R3 was expressed in a subset of taste receptor cells in taste buds and that the topographic distribution of T1R3 in various taste papillae was different from those of the other T1R members. Genetic mapping of T1R3 with a mouse/hamster radiation hybrid panel located the gene on the distal end of mouse chromosome 4 correlated with the Sac locus affecting sweet sensitivity of mice. Our results indicate that T1R3 may serve as the receptor for sweet perception in mice.

Amino Acid Sequence↗
PubMed1988

Cyclic nucleotides may mediate taste transduction.

Taste stimulus adsorption is believed to occur at the taste cell microvillous membrane. But due to technical difficulties of inserting glass electrodes into the mammalian taste cell, little is known about the mechanisms of taste transduction. Reliable intracellular recordings are necessary to determine the characteristics of taste cells. This has been accomplished previously in the mouse and is reported here. Recent experiments indicated that cyclic nucleotides can act on the inner surface of the membranes of a variety of cells to alter their ion-channel activity, and these substances might act as intracellular transmitters in taste cells. But tight junctions found at the apical membrane of mammalian taste cells do not allow stimuli to enter the taste bud, making it difficult to alter the environment of the taste cell by perfusing with chemical solutions. Here we report that cyclic AMP, cyclic GMP, EGTA or tetraethyl-ammonium electrophoretically injected into the mouse taste cell induce membrane depolarization and increased membrane resistance. These results suggest that a cyclic nucleotide enzymatic cascade, modulated by calcium ions, may mediate the potassium permeability that controls taste, in a way analogous to visual and olfactory transduction.

Animals↗
PubMed1964

THE RECEPTOR POTENTIAL OF THE TASTE CELL OF THE RAT.

The electrical responses of the taste cell of the rat to chemical stimuli were studied by means of microelectrode techniques. Although large positive potential changes in the taste cell were usually elicited by taste stimuli, the response was a small negative potential change with respect to surrounding tissues if the microelectrode was thrust deeply into the taste bud. Both FeCl(3) and cocaine produced a positive change in the steady potential. If this new potential is larger than a certain equilibrium potential, reversal of the polarity of the potential change caused by a taste stimulus is observed. Gamma-aminobutyric acid and acetylcholine had no effect on the receptor steady potential nor on the receptor responses elicited by taste stimuli.

Cocaine↗
PubMed2004

Morphologic characterization of rat taste receptor cells that express components of the phospholipase C signaling pathway.

Rat taste buds contain three morphologically distinct cell types that are candidates for taste transduction. The physiologic roles of these cells are, however, not clear. Inositol 1,4,5-triphosphate (IP(3)) has been implicated as an important second messenger in bitter, sweet, and umami taste transductions. Previously, we identified the type III IP(3) receptor (IP(3)R3) as the dominant isoform in taste receptor cells. In addition, a recent study showed that phospholipase Cbeta(2) (PLCbeta(2)) is essential for the transduction of bitter, sweet, and umami stimuli. IP(3)R3 and PLCbeta(2) are expressed in the same subset of cells. To identify the taste cell types that express proteins involved in PLC signal transduction, we used 3,3'diaminobenzidine tetrahydrochloride immunoelectron microscopy and fluorescence microscopy to identify cells with IP(3)R3. Confocal microscopy was used to compare IP(3)R3 or PLCbeta(2) immunoreactivity with that of some known cell type markers such as serotonin, protein gene-regulated product 9.5, and neural cell adhesion molecule. Here we show that a large subset of type II cells and a small subset of type III cells display IP(3)R3 immunoreactivity within their cytoplasm. These data suggest that type II cells are the principal transducers of bitter, sweet, and umami taste transduction. However, we did not observe synapses between type II taste cells and nerve fibers. Interestingly, we observed subsurface cisternae of smooth endoplasmic reticulum at the close appositions between the plasma membrane of type II taste cells and nerve processes. We speculate that some type II cells may communicate to the nervous system via subsurface cisternae of smooth endoplasmic reticulum in lieu of conventional synapses.

Animals↗
PubMed1999

Localisation of actin, villin, fimbrin, ezrin and ankyrin in rat taste receptor cells.

Mammalian taste buds consist of 50-150 pear-or spindle-shaped taste receptor cells which contain, at their apical cell surface, a bundle of microvillar projections. The microvilli probably serve to increase the receptive membrane surface of the chemosensory receptor cells. The molecular basis controlling the ultrastructure of taste receptor microvilli is present unknown. In the present study we analysed, by immunostaining at the light and electron microscopic levels and by immunoblotting, components of the cytoskeleton of these microvilli. We show here that taste cell microvilli contain the major cytoskeletal proteins of intestinal microvilli, actin, fimbrin and villin. Another actin-binding, peripheral membrane protein of intestinal microvilli, ezrin, was also localised to taste cell microvilli, where ezrin might play a role, for example, in placement of specific membrane proteins to the microvillus membrane. In search of further linkage proteins, we found ankyrin localised along the basolateral cell surface of taste receptor cells, where ankyrin might be involved in the immobilisation of the Na+, K+-ATPase or other ion-translocating proteins of taste cells to the membrane cytoskeleton.

Actins↗
PubMed1967

Altered temperature and taste responses from cross-regenerated sensory nerves in the rat's tongue.

1. The chorda tympani nerve, which innervates the front of the rat tongue, was found to be much less responsive to tongue cooling than the IXth nerve, which innervates the back of the tongue. The two nerves also differed in their relative responsiveness to various taste chemicals.2. Through cross-union the IXth nerve was made to innervate the front of the tongue, and in other rats the chorda tympani nerve the back of the tongue.3. After an average of 15 post-operative weeks, electrophysiological recordings of whole nerve action potential discharges were made from normal, control regenerated, and cross-regenerated nerves. Cooling, and chemical and mechanical stimulation of the tongue demonstrated that the control regenerated and cross-regenerated nerves had established functional connexions.4. Neither the response to cooling nor the relative taste responses were altered by either of two types of control chorda tympani nerve regeneration.5. In contrast, the cross-regenerated chorda increased its responsiveness to tongue cooling and the cross-regenerated IXthe nerve lost much of its responsiveness to cooling.6. Cross-regeneration also caused the relative taste responses to change and appear quite similar to the responses obtained from the nerve which normally innervated that tongue region (e.g. the cross-regenerated IXth nerve responded like a chorda tympani nerve).7. It is suggested that the sensory response evoked in the chorda tympani and IXth nerves by tongue cooling or taste stimulation is at least partially dependent upon the character of the tongue tissue in which the nerve terminates-the epithelium at the front differs from that at the back of the rat tongue.8. These results rule out the following two hypotheses: (a) that the nerve ending itself functions as a taste receptor in direct contact with applied chemicals and yet is uninfluenced by the character of the tissue in which it terminates, (b) that assuming the taste bud cells are an integral part of the receptive process, the taste nerve ending determines the chemical specificity of the taste cell which it induces, without any previous modification of the nerve by the tissue in which it terminates.

Action Potentials↗
PubMed1993

Fluorescent dye (DiI) reveals the sensory cells in the lingual epithelium: a confocal laser scanning microscopic study.

Fluorescent carbocyanine dye was applied to the glossopharyngeal nerve of axolotls fixed with paraformaldehyde. Three cell types in the lingual epithelium were transneuronally labeled: chemosensory taste receptor cells and presumably mechanosensory basal cells in the taste buds, and epithelial cells with unknown function in the non-taste lingual epithelium.

Ambystoma mexicanum↗
PubMed1965

DETERGENTS: EFFECTS ON THE CHEMICAL SENSES OF THE FISH ICTALURUS NATALIS (LE SUEUR).

Two types of detergents, one easily oxidized and the other not, damage the chemoreceptors of yellow bullheads, Ictalurus natalis (le Sueur) exposed to concentrations of 0.5 parts per million in the surrounding water. This concentration is considerably lower than that at which sublethal damage has been detected previously. Histological examination reveals erosion of the taste buds; electrophysiological methods and observations of swimming and feeding behavior reveal impairment of receptor function. Affected fish do not fully recover after 6 weeks in detergent-free water.

Animals↗
PubMed2006

Taste responsiveness of fungiform taste cells with action potentials.

It is known that a subset of taste cells generate action potentials in response to taste stimuli. However, responsiveness of these cells to particular tastants remains unknown. In the present study, by using a newly developed extracellular recording technique, we recorded action potentials from the basolateral membrane of single receptor cells in response to taste stimuli applied apically to taste buds isolated from mouse fungiform papillae. By this method, we examined taste-cell responses to stimuli representing the four basic taste qualities (NaCl, Na saccharin, HCl, and quinine-HCl). Of 72 cells responding to taste stimuli, 48 (67%) responded to one, 22 (30%) to two, and 2 (3%) to three of four taste stimuli. The entropy value presenting the breadth of responsiveness was 0.158 +/- 0.234 (mean +/- SD), which was close to that for the nerve fibers (0.183 +/- 0.262). In addition, the proportion of taste cells predominantly sensitive to each of the four taste stimuli, and the grouping of taste cells based on hierarchical cluster analysis, were comparable with those of chorda tympani (CT) fibers. The occurrence of each class of taste cells with different taste responsiveness to the four taste stimuli was not significantly different from that of CT fibers except for classes with broad taste responsiveness. These results suggest that information derived from taste cells generating action potentials may provide the major component of taste information that is transmitted to gustatory nerve fibers.

Action Potentials↗
PubMed2001

A brief survey of the modifications in sensory-secretory organs of the neonatal rat tongue.

Recent data obtained on rats suggest that in the days immediately following birth several events take place in the circumvallate papillae of the oral cavity. A phylogenetically primitive system of solitary chemosensory cells develops and is rapidly replaced by taste buds. The lipase-secreting von Ebner gland, which is associated with taste organs, begins to develop by forming short tubules. The intrinsic nervous system of the gustatory organs rapidly completes its maturation showing fast proliferation of fibers and immunocytochemical maturation. Intraepithelial lipid accumulation is visible in the non-receptorial mucosa of the tongue, showing aspects which suggest an active lipid secretion. These data demonstrate that in the rat the structure of the sensory-secretory organs of the newborn's tongue shows a typical conformation with respect to the adult and rapidly changes its organization in the first week after the birth. At the present level of knowledge, it is difficult to link the anatomical structures to peculiar functional roles but the rather simple organization of the neonatal gustatory epithelium could be in relation to the dietary regimen. The data obtained in laboratory animals underline the necessity of studies on human newborns to update the anatomical knowledge of the oral chemoceptive system.

Animals↗
PubMed2004

Support of trigeminal sensory neurons by nonneuronal p75 neurotrophin receptors.

The p75 neurotrophin receptor (p75NTR) binds all four mammalian neurotrophins, including neurotrophin-3 (NT-3) required for the development of select sensory neurons. This study demonstrated that many gustatory and somatosensory neurons of the tongue depend upon p75NTR. Each of thousands of filiform papillae at the front of the tongue as well as each somatosensory prominence at the back of the tongue has a small cluster of p75NTR-positive epithelial cells that is targeted by somatosensory innervation. This expression of p75NTR by epithelial target cells required NT-3 but not adult innervation. NT-3-secreting cells were adjacent to the p75NTR-positive target cells of each somatosensory organ, as demonstrated in NT-3(lacZneo) transgenic mice. In NT-3 null mutant mice, there were few lingual somatosensory neurons. In p75NTR null mutant mice, the lingual somatosensory axons were likewise absent or had deficient terminal arborizations. Cell culture indicated that substrate p75NTR can influence neuronal outgrowth. Specifically, dissociated trigeminal sensory neurons more than doubled their neurite lengths when grown on a lawn of p75NTR-overexpressing fibroblasts. This enhancement of neurite outgrowth by fibroblast p75NTR raises the possibility that epithelial target cell p75NTR may help to promote axonal arborization in vivo. The co-occurrence in p75NTR null mice of a 35% reduction in geniculate ganglion taste neurons and a shortfall of taste buds is consistent with the established role of gustatory innervation in prompting mammalian taste receptor cell differentiation.

Animals↗
PubMed2000

Inhibitory effect of gurmarin on palatal taste responses to amino acids in the rat.

Gurmarin (10 microg/ml), a protein extracted from Gymnema sylvestre, depressed significantly (40-50%) the phasic taste responses to sugars (sucrose, fructose, lactose, and maltose) and saccharin sodium recorded from the greater superficial petrosal nerve (GSP) innervating palatal taste buds in the rat. However, no significant effect of gurmarin was observed for taste responses to NaCl, HCl, and quinine hydrochloride. Phasic responses to D-amino acids that taste sweet to humans (His, Asn, Phe, Gln) were also depressed, but gurmarin treatment was without significant effect on taste responses to D-Trp and D-Ala, six L-amino acids (His, Asn, Phe, Gln, Trp, and Ala), and two basic amino acid HCl salts (Arg and Lys). With the exception of D-Trp, these inhibitory effects of gurmarin on GSP taste responses were related to the rat's preference for these substances.

Amino Acids↗
PubMed2006

Umami responses in mouse taste cells indicate more than one receptor.

A number of gustatory receptors have been proposed to underlie umami, the taste of L-glutamate, and certain other amino acids and nucleotides. However, the response profiles of these cloned receptors have not been validated against responses recorded from taste receptor cells that are the native detectors of umami taste. We investigated umami taste responses in mouse circumvallate taste buds in an intact slice preparation, using confocal calcium imaging. Approximately 5% of taste cells selectively responded to L-glutamate when it was focally applied to the apical chemosensitive tips of receptor cells. The concentration-response range for L-glutamate fell approximately within the physiologically relevant range for taste behavior in mice, namely 10 mm and above. Inosine monophosphate enhanced taste cell responses to L-glutamate, a characteristic feature of umami taste. Using pharmacological agents, ion substitution, and immunostaining, we showed that intracellular pathways downstream of receptor activation involve phospholipase C beta2. Each of the above features matches those predicted by studies of cloned and expressed receptors. However, the ligand specificity of each of the proposed umami receptors [taste metabotropic glutamate receptor 4, truncated metabotropic glutamate receptor 1, or taste receptor 1 (T1R1) and T1R3 dimers], taken alone, did not appear to explain the taste responses observed in mouse taste cells. Furthermore, umami responses were still observed in mutant mice lacking T1R3. A full explanation of umami taste transduction may involve novel combinations of the proposed receptors and/or as-yet-undiscovered taste receptors.

Animals↗
PubMed1992

Responses of single facial taste fibers in the channel catfish, Ictalurus punctatus, to amino acids.

1. Amino acids and nucleotides stimulate taste receptors of teleosts. In this report, responses to these compounds of 105 facial taste fibers (79 fully characterized) that innervate maxillary barbel taste buds of the channel catfish (Ictalurus punctatus) were analyzed. 2. The fully characterized facial taste fibers that responded to amino acids (n = 68) were generally poorly responsive to nucleotides and related substances (NRS), whereas the fibers responsive to NRS (n = 11) were poorly responsive to amino acids. Spike discharge of the amino acid-responsive fibers to the most potent amino acid stimulus tested per fiber increased 44-fold from a mean spontaneous activity of 2.1 +/- 3.5 to 92.1 +/- 42.4 (SD) spikes/3 s. Spike activity of the NRS-responsive fibers to NRS increased 11.5-fold from a mean spontaneous activity of 3.4 +/- 5.9 to 39.1 +/- 27.4 spikes/3 s. There was no significant difference between the spontaneous rates, but stimulus evoked spike rates for the amino acid-responsive fibers were significantly greater (P < 0.05; Mann-Whitney test) than those for the NRS-responsive fibers. 3. Hierarchical cluster analysis based on the 3-s response time identified three major groups of neurons. The identified clusters comprised neurons that were highly responsive to either L-alanine (i.e., Ala cluster; n = 39), L-arginine (i.e., Arg cluster; n = 29), or NRS (NRS cluster; n = 11). Fibers comprising the Arg cluster were more narrowly tuned than those within the Ala cluster. This report further characterizes the responses to amino acids of the individual facial taste fibers comprising the Ala and Arg clusters. 4. Subclusters were evident within both of the amino acid-responsive clusters. The Arg cluster was divisible into two subclusters dependent on the response to 1 mM L-proline. Twelve neurons that were significantly (P < 0.05; Mann-Whitney test) more responsive to L-proline than the remaining 17 neurons within the Arg cluster formed the Arg/Pro subcluster; these latter 17 neurons comprised the Arg subcluster. However, there was no significant difference (Mann-Whitney test) in the response to L-arginine between fibers within either subcluster across four different response times analyzed. Fibers within the Ala cluster were generally poorly responsive to L-proline. Four alanine subclusters were suggested on the basis of their relative responses to L-alanine, D-alanine, L-arginine, and the NRS; however, of the 39 fibers comprising the alanine cluster, two alanine subclusters comprised only two fibers each, and the third subcluster consisted of four fibers.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed2000

Peripheral glutamate receptors: molecular biology and role in taste sensation.

Glutamate is the most widespread excitatory neurotransmitter in the mammalian brain. Two classes of glutamate receptor have been cloned, the ionotropic (ligand-gated ion channels) and the metabotropic (G protein-coupled receptors). Three subclasses of ionotropic glutamate receptors are known; they are named after selective agonists, i.e., alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA), N-methyl-D-aspartate (NMDA) and kainate receptors. Fifteen functional subunits assemble together in heteromultimeric complexes to form these receptors as follows: GluR1-GluR4 for AMPA; GluR5-GluR7 and KA1-KA2 for kainate; and NR1, NR2A-NR2D and NR3 for NMDA receptors. Within a subclass, the subunit composition strongly influences the pharmacologic and biophysical properties of the receptors. The metabotropic glutamate receptors fall into the following three groups, each containing two or more individual receptor proteins: group I (mGluR1, mGluR5), group II (mGluR2, mGluR3), and group III (mGluR4, mGluR6, mGluR7 and mGluR8). In contrast to the ionotropic receptors, the metabotropic glutamate receptors appear to act as monomers or homodimers rather than heteromers. Messenger RNAs encoding several ionotropic subunits and a mGluR4-like receptor have been identified in taste buds. Although controversial, the evidence is consistent with an NMDA receptor serving as a primary taste transducer for monosodium glutamate (MSG), and a metabotropic glutamate receptor modulating the flavor-enhancing effect of MSG. Thus the neurotransmitter glutamate is intimately involved in the central processing of taste information.

Animals↗
PubMed1998

Self-inhibition in amiloride-sensitive sodium channels in taste receptor cells.

Electrophysiological recording techniques were used to study the Na+ dependence of currents through amiloride-sensitive sodium channels (ASSCs) in rat taste cells from the fungiform and vallate papillae. Perforated patch voltage clamp recordings were made from isolated fungiform and vallate taste receptor cells (TRCs) and Na+ transport was measured across lingual epithelia containing fungiform or vallate taste buds in a modified Ussing chamber. In isolated fungiform TRCs that contain Na+ currents sensitive to the diuretic amiloride, Na+ ions inhibit their own influx through ASSCs, a process known as sodium self-inhibition. Due to the interaction between self-inhibition and the driving force for Na+ entry, self-inhibition is most evident in whole-cell recordings at Na+ concentrations from 50 to 75 mM. In amiloride-sensitive cells, the Na permeability is significantly higher in extracellular solutions containing 35 mM Na+ than in 70 or 140 mM Na+. Compared with the block by amiloride, the development of self-inhibition is slow, taking up to 15 s to become maximally inhibited. Approximately one third of fungiform TRCs and all vallate TRCs lack functional ASSCs. These amiloride-insensitive TRCs show no signs of self-inhibition, tying this phenomenon to the presence of ASSCs. The sulfhydryl reagent, p-hydroxymercuribenzoate (p-HMB; 200 microM), reversibly removed self-inhibition from amiloride-sensitive Na+ currents, apparently by modifying cysteine residues in the ASSC. Na+ currents in amiloride-insensitive TRCs were unaffected by p-HMB. In sodium transport studies in fungiform taste bud-containing lingual epithelia, approximately 40% of the change in short-circuit current (Isc) after addition of 500 mM NaCl to the mucosal chamber is amiloride sensitive (0.5 mM). p-HMB significantly enhanced mucosal NaCl-induced changes in these epithelia at mucosal Na+ concentrations of 50 mM and above. In contrast, the vallate-containing epithelia, which are insensitive to amiloride, showed no enhancement of Isc during p-HMB treatment. These findings suggest that sodium self-inhibition is present in ASSCs in taste receptor cells where it may play a crucial role in performance of salt-sensitive pathways in taste tissue during sodium stimulation. This phenomenon may be important in the process of TRC adaptation, in the conservation of cellular resources during chronic sodium exposure, or in the gustatory response to water.

Amiloride↗
PubMed1994

Human taste cells express the G protein alpha-gustducin and neuron-specific enolase.

Expression of the alpha-subunit of the taste-specific G protein alpha-gustducin and the glycolytic enzyme neuron-specific enolase (NSE) was investigated immunohistochemically in human circumvallate and foliate taste papillae. Immunofluorescence for alpha-gustducin was observed in taste cells of both types of papillae and exhibited two patterns of immunofluorescence, plasmalemmal and cytosolic. The plasmalemmal pattern showed intense immunofluorescence localized to the apical region, and was exhibited by most immunoreactive taste cells. In contrast, the cytosolic pattern, observed in one or two immunoreactive cells in a taste bud per section, showed immunofluorescence distributed throughout the cytoplasm. A subpopulation of alpha-gustducin-immunoreactive taste receptor cells, most of which exhibited the cytosolic pattern, also expressed NSE. Optical sectioning, using confocal laser scanning microscopy, demonstrated the highest level of expression of alpha-gustducin in the apical microvillar region of the taste cells in close apposition to the taste pore. These studies indicate conservation of epitopes of alpha-gustducin in humans and rats, and suggest that this G protein is associated with taste transduction in both rats and humans. The patterns of expression of alpha-gustducin, and coexpression with NSE, may correlate with specialized subtypes or developmental stages of taste receptor cells.

Aged↗
PubMed1996

Neonatal chorda tympani transection alters adult preference for ammonium chloride in the rat.

The immature gustatory system of the neonatal rat is characterized by sensitivity to disruption by early interventions such as receptor or nerve damage. The present studies examined the effect of chorda tympani transection (neoCTX) of neonates on adult preference for salt and nonsalt stimuli. NeoCTX at 10 days of age led to a striking change in adult rats' preference for NH4Cl solutions but little change in preference for other solutions, including NaCl and KCl. Permanent anatomical effects of neoCTX included failure of the nerve to regenerate and a loss of all fungiform taste buds. Preference for NH4Cl was not due to an inability to discriminate it from NaCl. Following taste aversion conditioning to NaCl, neoCTX rats clearly distinguished between NaCl and NH4Cl. The effects on NH4Cl preference reflect a sensitive period during development because adult rats receiving similar surgery did not show any change in NH4Cl preference.

Ammonium Chloride↗