David V. Smith

Neural coding of gustatory information.

The nervous system encodes information relating chemical stimuli to taste perception, beginning with transduction mechanisms at the receptor and ending in the representation of stimulus attributes by the activity of neurons in the brain. Recent studies have rekindled the long-standing debate about whether taste information is coded by the pattern of activity across afferent neurons or by specifically tuned labeled lines. Taste neurons are broadly tuned to stimuli representing different qualities and are also responsive to stimulus intensity and often to touch and temperature. Their responsiveness is also modulated by a number of physiological factors. In addition to representing stimulus quality and intensity, activity in taste neurons must code information about the hedonic value of gustatory stimuli. These considerations suggest that individual gustatory neurons contribute to the coding of more than one stimulus parameter, making the response of any one cell meaningful only in the context of the activity of its neighbors.

GABA-mediated corticofugal inhibition of taste-responsive neurons in the nucleus of the solitary tract.

The nucleus of the solitary tract (NST) receives descending connections from several forebrain targets of the gustatory system, including the insular cortex. Many taste-responsive cells in the NST are inhibited by gamma-aminobutyric acid (GABA). In the present study, we investigated the effects of cortical stimulation on the activity of gustatory neurons in the NST. Multibarrel glass micropipettes were used to record the activity of NST neurons extracellularly and to apply the GABA(A) antagonist bicuculline methiodide (BICM) into the vicinity of the cell. Taste stimuli were 0.032 M sucrose (S), 0.032 M NaCl (N), 0.00032 M citric acid (H), and 0.032 M quinine hydrochloride (Q), presented to the anterior tongue. Each of 50 NST cells was classified as S-, N-, H-, or Q-best on the basis of its response to chemical stimulation of the tongue. The ipsilateral insular cortex was stimulated both electrically (0.5 mA, 100 Hz, 0.2 ms) and chemically (10 mM DL-homocysteic acid, DLH), while the spontaneous activity of each NST cell was recorded. The baseline activity of 34% of the cells (n=17) was modulated by cortical stimulation: eight cells were inhibited and nine were excited. BICM microinjected into the NST blocked the cortical-induced inhibition but had no effect on the excitatory response. Although the excitatory effects were distributed across S-, N-, and H-best neurons, the inhibitory effects of cortical stimulation were significantly more common in N-best cells. These data suggest that corticofugal input to the NST may differentially inhibit gustatory afferent activity through GABAergic mechanisms.

Taste quality profiles for fifteen organic and inorganic salts

Biophysical studies of isolated taste receptor cells show that one transduction mechanism for Na+ salts involves the inward movement of Na+ through an apical ion channel, which is sensitive to the diuretic amiloride. An additional paracellular pathway also appears to be involved in NaCl transduction, but not in the transduction of organic Na+ salts. Little is known, however, about how these receptor mechanisms relate to taste perception. Recent human psychophysical studies suggest that the amiloride-sensitive transduction pathway is coupled to the sour side taste of these salts rather than to their saltiness. In the present study, we employed direct magnitude estimation of taste intensity and quality of fifteen organic and inorganic Na+, Li-, K+, and Ca+2 salts. Many salts had multiple taste qualities, such as the salty and bitter tastes of NH4Cl and KCl; the Ca+2 salts were predominantly bitter. Taste quality often changed with stimulus concentration. Multivariate analyses of their taste profiles resulted in a grouping of these 18 stimuli within a taste space bounded by NaCl, sucrose, citric acid, and QHCl, with the organic salts positioned between NaCl and citric acid. The organic Na+ salts and the Li+ salts were considerably less salty and proportionately more sour than NaCl. These results, combined with previous work showing that amiloride suppresses the sourness of NaCl and Na-gluconate, predict that the organic Na+ salts and the Li+ salts would be more greatly suppressed by amiloride treatment than would NaCl.

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.

Amiloride suppression of the taste intensity of sodium chloride: evidence from direct magnitude scaling.

The transduction of Na+ salts has been shown in many species to be mediated in part by an epithelial ion channel on the apical surface of the taste receptor cell membrane, which is blockable by the diuretic amiloride. In addition to this apical ion channel, Na+ is also transduced via a paracellular pathway, which is not sensitive to amiloride. There are significant species differences in the contribution of the amiloride-sensitive pathway to Na+ transduction. Previous human psychophysical studies have reported conflicting results on the effects of amiloride in suppressing the intensity of NaCl. In general, these studies used amiloride doses that were much higher than those showing clear suppressive effects in electrophysiological studies in other species. In the present experiment, we used direct magnitude scaling of the intensities of five NaCl concentrations flowed over the anterior portion of the tongue to determine the effects of amiloride treatment at lower doses. NaCl was presented after adaptation of the tongue to water or mixed with and presented after adaptation to 10, 50, or 100 microM amiloride-HCl. Subjects estimated the intensity of NaCl and of these concentrations of amiloride in each treatment condition using magnitude estimation with a 0.1 M NaCl modulus presented following a water rinse prior to each session. Results showed that amiloride had a significant suppressive effect on the perceived intensity of NaCl, with a similar effect seen at all three amiloride doses. The psychophysical function after amiloride showed a parallel shift to the right. The average suppression over all NaCl concentrations was 21%.(ABSTRACT TRUNCATED AT 250 WORDS)

Opioid modulation of taste responses in the nucleus of the solitary tract.

Gustatory processing within the medulla is modulated by a number of physiologic and experiential factors. Several neurotransmitters, including excitatory amino acids, GABA, and substance P, are involved in synaptic processing within the rostral portion of the nucleus of the solitary tract (NST). Endogenous opiates have been implicated in the regulation of feeding behavior and in taste palatability and gustatory responses in the parabrachial nuclei are reduced by systemic morphine. In the present experiments, extracellular recording of neuronal activity within the NST in response to taste input was combined with local microinjection of met-enkephalin (Met-ENK) and naltrexone (NLTX) to determine the effect of these agents on gustatory activity. The anterior tongue was stimulated with anodal current pulses to determine the time course of drug action (n=85 cells) and with prototypical taste stimuli (0.032 M sucrose, NaCl, and quinine hydrochloride, and 0.0032 M citric acid) to investigate the effects of these opioid compounds on taste-evoked responses (n=80 cells). Among these 165 taste-responsive neurons in the NST, the activity of 39 (23.6%) was suppressed by Met-ENK. These effects were dose-dependent and blockable by NLTX, which alone was without effect, suggesting that opiates do not maintain a tonic inhibitory influence. Immunohistochemical experiments demonstrated both micro - and delta-opioid receptors within the gustatory portion of the NST; previous studies had shown numerous fiber terminals containing Met-ENK. These data suggest that endogenous opiates play an inhibitory role in gustatory processing within the medulla.

Neural coding of aversive and appetitive gustatory stimuli : interactions in the hamster brain stem

There is increasing evidence, both electrophysiological and behavioral, that bitter and sweet stimuli drive parallel pathways in the gustatory brainstem. Here we report two lines of investigation that suggest significant interactions among these parallel systems. First, responses recorded from single cells in the hamsters parabrachial nuclei (PbN) show that quinine hydrochloride (QHCl) produces a substantial suppression (> 40%) of the responses of PbN cells to sucrose. Sucrose stimulation has a reciprocal suppressive effect on the response to QHCl. These results imply that aversive and appetitive stimuli produce mutual inhibition in the gustatory system; studies of the chorda tympani nerve response suggest that this inhibition likely arises within the brainstem. A second line of investigation, using both an in vitro brainstem slice preparation and in vivo pharmacological manipulations of cells in the hamster NST, has demonstrated an inhibitory network within the rostral NST that plays a role in the modulation of taste activity. Patch-clamp and extracellular recording studies in vitro show that cells within the rostral central subdivision of the NST are inhibited by gamma-aminobutyric acid (GABA); this mediation is largely through the GABAA receptor subtype. Here we show that responses to taste stimulation recorded extracellularly from NST cells in vivo can be inhibited by local microinjections of GABA; this inhibition is blocked by the GABAA receptor antagonist bicuculline methiodide. Responses to sucrose are significantly more inhibited than those to NaCl or KCl. These combined lines of evidence show that appetitive and aversive stimuli activate mutually inhibitory systems within the brainstem and suggest that the basis for this interaction is a GABAergic inhibitory network within the NST.(ABSTRACT TRUNCATED AT 250 WORDS)

Distribution and synaptology of glossopharyngeal afferent nerve terminals in the nucleus of the solitary tract of the hamster.

The distribution and synaptology of the afferent fibers of the glossopharyngeal nerve (IXN) in the hamster were studied by using horseradish peroxidase (HRP) histochemistry visualized with light and electron microscopy. Crystals of HRP were applied to the trunk of IXN in the vicinity of the petrosal ganglion. The densest IXN afferent label was distributed within the nucleus of the solitary tract (nst), just caudal to but overlapping with the area of termination of the facial nerve. Labeled IXN fibers extended rostrally to the principal trigeminal nucleus and caudally to the cervical spinal cord. There was significant labeling within the spinal trigeminal complex; the area postrema and the medullary reticular formation contained some labeled fibers.

Amiloride suppresses the sourness of NaCl and LiCl.

The transduction of Na+ salts in many species is mediated by both apical and submucosal ion channels on the taste receptor-cell membrane. The apical ion channel is blockable by the diuretic amiloride, whereas the submucosal pathway is not. Previous human psychophysical studies have shown a decrease in NaCl taste intensity caused by amiloride that is smaller than the reduction of the electrophysiological response produced by amiloride in other species. Many salts, including NaCl, elicit not only a salty taste to humans, but also sweet, sour, or bitter side tastes. Amiloride has been shown to reduce the sourness, but not the saltiness, of NaCl and Na gluconate and to have no effect on the taste of KCl. The present experiment further evaluated the hypothesis that the sour taste of Na+ and Li+ salts is mediated by the amiloride-sensitive transduction mechanism, by examining the effect of amiloride on the taste of LiCl, which is considerably more sour than NaCl. Four concentrations of NaCl, LiCl, and KCl were presented to the anterior tongue following adaptation to water or after 10 microM amiloride treatment. Subjects estimated the intensity of the taste of each stimulus and divided this estimate among the appropriate taste qualities. There was a significant decrease in the total taste intensity of NaCl and LiCl after amiloride, but no effect on KCl; LiCl was more greatly suppressed than NaCl. These data show no effect on the saltiness of LiCl or NaCl, except for a small reduction in the saltiness of 0.1 M NaCl. On the contrary, there was a significant effect on the sourness of both NaCl and LiCl. Citric acid (3.2 mM) was also used as a stimulus, but amiloride treatment had no effect on its sourness. These data indicate that the amiloride-sensitive channel plays a key role in the perception of the sour taste of NaCl and LiCl (but not citric acid) and little role in the perception of saltiness. The salty taste of these salts may arise from other transduction pathways.

Differential expression of carbohydrate blood‐group antigens on rat taste‐bud cells: Relation to the functional marker α‐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 Lewisb, A, and H type 2 in taste buds of the rat oral cavity. We compared the distributions of these antigens with that of α‐gustducin, a G‐protein subunit implicated in responses to sweet‐ and bitter‐tasting substances. The A and Lewisb 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 Lewisb was not found elsewhere in the digestive tract. Lewisb and A were not removed by lipid extraction, suggesting that they are present on glycoproteins rather than glycolipids. All Lewisb‐positive cells expressed α‐gustducin, but only a fraction of α‐gustducin–positive cells expressed Lewisb. The fraction of taste‐bud cells expressing Lewisb decreased in the order: vallate papillae > foliate papillae > nasoincisor duct. The epiglottis had almost no taste‐bud cells that expressed Lewisb. The A antigen appeared on taste‐bud cells that also expressed α‐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 α‐gustducin in foliate and vallate papillae. In vallate papillae, cells expressed either A or Lewisb, but not both. Lewisb appears to be restricted to differentiated light cells that also express α‐gustducin and may be involved in intercellular interactions of these cells. J. Comp. Neurol. 415:230–239, 1999.