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J D Boughter

Publications and source records attributed to J D Boughter.

11 recordsLinked to original sources
PubMed2001

Distribution of gustatory sensitivities in rat taste cells: whole-cell responses to apical chemical stimulation.

Several taste transduction mechanisms have been demonstrated in mammals, but little is known about their distribution within and across receptor cells. We recorded whole-cell responses of 120 taste cells of the rat fungiform papillae and soft palate maintained within the intact epithelium in a modified Ussing chamber, which allowed us to flow tastants across the apical membrane while monitoring the activity of the cell with a patch pipette. Taste stimuli were: 0.1 m sucrose, KCl, and NH(4)Cl, 0.032 m NaCl, and 3.2 mm HCl and quinine hydrochloride (QHCl). When cells were held at their resting potentials, taste stimulation resulted in conductance changes; reversible currents >5 pA were considered reliable responses. Sucrose and QHCl produced a decrease in outward current and membrane conductance, whereas NaCl, KCl, NH(4)Cl, and HCl elicited inward currents accompanied by increased conductance. Combinations of responses to pairs of the four basic stimuli (sucrose, NaCl, HCl, and QHCl) across the 71-84 cells tested with each pair were predictable from the probabilities of responses to individual stimuli, indicating an independent distribution of sensitivities. Of 62 cells tested with all four basic stimuli, 59 responded to at least one of the stimuli; 16 of these (27.1%) responded to only one, 20 (33.9%) to two, 15 (25.4%) to three, and 8 (13.6%) to all of the basic stimuli. Cells with both inward (Na(+)) and outward (K(+)) voltage-activated currents were significantly more broadly tuned to gustatory stimuli than those with only inward currents.

Ammonium Chloride↗
PubMed1999

Differential expression of carbohydrate blood-group antigens on rat taste-bud cells: relation to the functional marker alpha-gustducin.

An afferent nerve fiber supplying a taste bud receives input from several taste receptor cells, yet is predominantly responsive to one of the classic taste qualities (salt, acid, sweet, or bitter). This specificity requires recognition between taste receptor cells and nerve fibers that may be mediated by surface markers correlating with function. In an effort to identify potential markers, we used immunofluorescence and confocal microscopy to examine expression of the oligosaccharide blood-group antigens Lewis(b), A, and H type 2 in taste buds of the rat oral cavity. We compared the distributions of these antigens with that of alpha-gustducin, a G-protein subunit implicated in responses to sweet- and bitter-tasting substances. The A and Lewis(b) antigens were present only on spindle-shaped cells whose apical processes reached the taste pore. These antigens were not present on epithelial cells surrounding taste buds, and Lewis(b) was not found elsewhere in the digestive tract. Lewis(b) and A were not removed by lipid extraction, suggesting that they are present on glycoproteins rather than glycolipids. All Lewis(b)-positive cells expressed alpha-gustducin, but only a fraction of alpha-gustducin-positive cells expressed Lewis(b). The fraction of taste-bud cells expressing Lewis(b) decreased in the order: vallate papillae > foliate papillae > nasoincisor duct. The epiglottis had almost no taste-bud cells that expressed Lewis(b). The A antigen appeared on taste-bud cells that also expressed alpha-gustducin in the order: foliate and vallate papillae > nasoincisor duct and epiglottis > fungiform papillae. In addition, the A antigen was present on many cells that lacked alpha-gustducin in foliate and vallate papillae. In vallate papillae, cells expressed either A or Lewis(b), but not both. Lewis(b) appears to be restricted to differentiated light cells that also express alpha-gustducin and may be involved in intercellular interactions of these cells.

Animals↗
PubMed1999

Cellular expression of alpha-gustducin and the A blood group antigen in rat fungiform taste buds cross-reinnervated by the IXth nerve.

Although taste buds are trophically dependent on their innervation, cross-reinnervation experiments have shown that their gustatory sensitivities are determined by the local epithelium. Both the gustatory G-protein, alpha-gustducin, and the cell-surface carbohydrate, the A blood group antigen, are expressed by significantly fewer fungiform than vallate taste cells in the rat. In these experiments, one side of the anterior portion of the tongue was cross-reinnervated by the IXth nerve in order to determine whether the molecular expression of taste bud cells is determined by the epithelium from which they arise or by the nerve on which they are trophically dependent. The proximal portion of the IXth nerve was anastomosed to the distal portion of the chorda tympani (CT) nerve using fibrin glue (IX-CT rats). Control animals had the CT cut and reanastomosed using the same technique (CT-CT rats), or had the CT avulsed from the bulla and resected to prevent regeneration (CTX rats). The animals survived for 12 weeks postoperatively, and the tongues were removed, stained with methylene blue, and the fungiform taste pores counted on both sides. Tissue from the anterior 5 mm of the tongue was cut into 50-microm sections, which were incubated with antibodies against alpha-gustducin and the human blood group A antigen. In both CT-CT and IX-CT rats, there was regeneration of fungiform taste buds, although in both groups there were significantly fewer taste buds on the operated side of the tongue. The normal vallate papilla had a mean of 8.37 alpha-gustducin-expressing cells and 5.22 A-expressing cells per taste bud, whereas the fungiform papillae contained 3.06 and 0.23 cells per taste bud, respectively. In both CT-CT and IX-CT rats there was a normal number of cells expressing alpha-gustducin or the A antigen in regenerated taste buds; in the CTX animals there was a significant decrease in the expression of these markers. These results demonstrate that the molecular phenotype of taste bud cells is determined by the local epithelium from which they arise and not by properties of the innervating nerve.

ABO Blood-Group System↗
PubMed1999

Neural representation of the taste of NaCl and KCl in gustatory neurons of the hamster solitary nucleus.

NaCl and KCl are monovalent salts that can be discriminated behaviorally by hamsters on the basis of their tastes. We examined the effects of the passive Na+ channel blocker amiloride on responses to both of these salts in 34 taste-responsive neurons of the nucleus of the solitary tract (NST) in the hamster. The effects of amiloride were assessed with two different, commonly employed stimulus protocols. Additionally, concentration-response functions for each salt were measured in 37 neurons. Cells were characterized by their best response to (in M) 0. 03 NaCl, 0.1 sucrose, 0.003 HCl, 0.001 quinine hydrochloride, and 0. 1 KCl. In neurons classified as NaCl-best, amiloride reversibly blocked responses to both NaCl and KCl. In neurons classified as HCl-best, amiloride had no effect on either stimulus. In sucrose-best neurons, amiloride blocked the response to NaCl but not KCl. These results support the hypothesis that both salts are transduced by at least two different receptor mechanisms. In the NST, information arising from these different inputs is maintained in discrete populations of neurons. In addition to differences in amiloride sensitivity, the cell types also differed in their responses to the salts across concentration. At midrange salt concentrations, NaCl-best neurons were far more responsive to NaCl than KCl, whereas HCl- and sucrose-best neurons responded equivalently to the two salts at all concentrations. Because NaCl- and HCl-best cells cannot by themselves distinguish NaCl from KCl, it is the relative activity across these cell types that comprises the code for taste discrimination.

Action Potentials↗
PubMed1998

Amiloride blocks acid responses in NaCl-best gustatory neurons of the hamster solitary nucleus.

Biophysical studies of isolated taste receptor cells show that one mechanism of Na+ salt transduction involves the inward movement of Na+ through amiloride-blockable ion channels on the apical receptor cell membrane, which leads to a direct depolarization. Hamster taste receptor cells with amiloride-blockable Na+ responses also show an amiloride-sensitive H+ current. Thus one mechanism for the transduction of acid taste involves the amiloride-sensitive channel. We investigated the effects of amiloride on responses to acids in neurons of the nucleus of the solitary tract (NST) of the hamster. The responses of 47 NST neurons were recorded extracellularly while the anterior tongue was stimulated with solutions representing the four taste qualities (NaCl, sucrose, HCl, quinine), which were used to characterize each cell on the basis of its best stimulus. The effects of amiloride on responses to 10 mM HCl, 10 mM citric acid, 100 mM NaCl, and 100 mM sucrose were then investigated. Stimuli were presented alone for 30 s (control trials) and also presented for 10 s, followed by a mixture of the stimulus with 10 microM amiloride for 10 s, followed by the stimulus alone again for 10 s (amiloride trials). The effects of amiloride were assessed by comparing the responses of cells with the stimulus + amiloride with that of the stimulus alone. In neurons classified as NaCl-best, amiloride reversibly blocked responses to NaCl, HCl, and citric acid. In HCl-best neurons, amiloride had no effect on responses to any of these stimuli. In sucrose-best neurons, amiloride blocked the response to NaCl but not to sucrose or to either acid. These results support the hypothesis that acids are transduced by at least two different receptor mechanisms in the hamster, amiloride sensitive and amiloride insensitive. At the NST, these inputs are tightly maintained in two separate populations of neurons. Sucrose-best neurons, which show amiloride effects on NaCl but not acids, appear to receive converging inputs from both amiloride-sensitive (N-best) and amiloride-insensitive (H-best) chorda tympani nerve fibers.

Action Potentials↗
PubMed1997

Behavioral specificity of the bitter taste gene Soa.

In mice, aversion to the bitter acetylated sugar sucrose octaacetate (SOA) is determined by a single genetic locus with three alleles. SWR/J (SW) inbred mice are SOA tasters: They avoid many compounds characterized as bitter-tasting by humans, at concentrations to which C3HeB/FeJ (C3:SOA demitasters) mice are less sensitive. C3.SW-Soa(a) congenic taster mice contain the taster allele transposed to a 99% C3 bitter-insensitive genetic background. SW, C3, C3.SW-Soa(a) congenic taster, and C3.SW demitaster mice were behaviorally tested with a series of 48-h two-bottle preference tests to determine the influence of the Soa(a) taster allele on sensitivity to a variety of bitter-tasting compounds. Soa allelic variation had a major effect on sensitivity to 0.003-1.0 mM SOA and several concentrations of the bitter-tasting alkaloids brucine, strychnine, and quinine. Effects were also found for 0.1 mM denatonium and 1 mM propylthiouracil. For caffeine, cycloheximide, thiamine, and two nonbitter compounds (NaCl and calcium hydroxide), the SW mice avoided lower concentrations than the other strains, but this avoidance was not due to the Soa(a) allele because both the C3 inbred and C3.SW-Soa(a) congenics were less sensitive. These results suggest the Soa gene product influences sensitivity to a subset of bitter-tasting compounds.

Alleles↗
PubMed1997

Differential expression of alpha-gustducin in taste bud populations of the rat and hamster.

The G-protein subunit alpha-gustducin, which is similar to rod transducin, has been implicated in the transduction of both sweet- and bitter-tasting substances. In rodents, there are differences in sensitivity to sweet and bitter stimuli in different populations of taste buds. Rat fungiform taste buds are more responsive to salts than to sweet stimuli, whereas those on the palate respond predominantly to sweet substances. In contrast, hamster fungiform taste buds are more sensitive to sweet-tasting stimuli. Taste buds in the vallate and foliate papillae of both species are sensitive to bitter compounds. These differences in sensitivity should be reflected in the numbers of gustducin-containing cells in different taste bud populations. We examined taste buds in the rat and hamster for immunoreactivity to an antibody against alpha-gustducin. Immunofluorescence of labeled taste cells was examined by confocal microscopy, and the cells were counted. Gustducin-positive cells were seen in all taste bud regions; they were spindle-shaped, with circular cross-sections and apical processes that extended to the taste pore. Cells with this characteristic shape in rat vallate taste buds are Type II (light) cells. In the rat, taste buds of the fungiform papillae had fewer gustducin-positive cells (3.1/taste bud) than those of other regions, including the posterior tongue and palate (>8.9/taste bud). Hamster fungiform taste buds contained twice as many gustducin-expressing cells (6.8/taste bud) as those of the rat. These data support the hypothesis that alpha-gustducin is involved in the transduction of both sweet- and bitter-tasting stimuli by mammalian taste receptor cells.

Animals↗
PubMed1995

C3.SW-Soaa heterozygous congenic taster mice.

Sensitivity to the bitter acetylated sugar sucrose octaacetate (SOA) is mediated by a single-locus system with three alleles in mice. Inbred strains are classified according to SOA phenotype as tasters, nontasters, or demitasters (intermediate sensitivity). A congenic quartet created from taster and nontaster strains has been used to investigate the effect of the Soa locus on non-SOA aspects of taste sensitivity. In this study, we created a third congenic set, C3.SW-Soaa, from taster (SWR/J) and demitaster (C3HeB/FeJ) strains. After 11 lineal backcross generations these C3.SW mice carry the tasting allele on a 99% C3 demitaster background. After testing a total of 938 mice, taster-demitaster proportions across 12 generations were consistent with expectations from a monogenic model. The resultant three-strain congenic series will allow further examination of the mechanisms of taste.

Alleles↗
PubMed1994

Hydrolysis of sucrose octa-acetate: qualitative differences in taster and demistaster avoidance phenotypes.

Calcium hydroxide and sodium hydroxide were used to hydrolyse sucrose octa-acetate (SOA) as a means of evaluating the taster (Soaa) and demitaster (Soac) allelic phenotypes of the genetic locus Soa. The SWR/J (taster) inbred strain and the B6.SW Soaa (taster) congenic strain were demonstrated to cease avoiding upon nearly complete hydrolysis of 10(-5) M SOA with calcium hydroxide or sodium hydroxide and of 10(-4) M SOA with calcium hydroxide. The BALB/cByJ, C3HeB/FeJ and DBA/2J (demitaster) inbred strains were demonstrated to cease avoiding after only a partial hydrolysis of 10(-3) M SOA using calcium hydroxide. It is suggested that specificity for the number or placement of the acetates of SOA underlies the difference between the taster and demitaster phenotypes.

Animals↗
PubMedDate not supplied

Neuronal cell types and taste quality coding.

Over the past 25 years, there have been two opposing views of how taste information is represented in the activity of gustatory neurons. One view, the across-fiber pattern (AFP) theory, postulates that taste quality is represented by the pattern of activity across the afferent population. Stimuli with similar tastes produce similar patterns of activity. The other view is that activity in a few distinct neuron types codes taste quality in a "labeled-line" fashion. Neurons responding best to sucrose, for example, would represent "sweetness," and those responding best to NaCl would code "saltiness." Some of these neuron types appear to have a biological significance, such as the NaCl-best cells, which receive input about sodium stimuli exclusively from an amiloride-sensitive epithelial ion channel. However, the relatively broad tuning of these neurons makes it unlikely that they are capable of unambiguously coding information about taste quality. Rather, these neuron types play a critical role in establishing unique AFPs that distinguish among taste stimuli. The relative activity across these cell types represent taste quality, much like the patterns of activity across broadly tuned photoreceptors code information about stimulus wavelength.

Animals↗