Thomas P. Hettinger

Specificity of amiloride inhibition of hamster taste responses.

Amiloride, a blocker of epithelial sodium channels, was found to have significant effects on electrophysiological and behavioral taste responses in the golden hamster (Mesocricetus auratus). Recordings from the whole chorda tympani nerve showed that amiloride rapidly, reversibly, and competitively inhibited responses to NaCl applied to the anterior tongue. The apparent dissociation constant for amiloride binding, extrapolated to zero NaCl concentration, was 10 nM, a value comparable to estimates for various transporting tight epithelia. Recordings from single chorda tympani nerve fibers showed that 10 microM amiloride completely inhibited responses of Na-selective N fibers but had minimal effect on responses of electrolyte-sensitive H fibers, even though both types of fibers responded well to 0.1 M NaCl. Sucrose responses were not affected by amiloride. Addition of 100 microM amiloride to 0.1 M NaCl consistently increased consumption of NaCl in two-bottle drinking tests. These data suggest that one mechanism by which the taste of NaCl is sensed, which does not require adsorption or a second messenger, involves entry of Na+ into taste bud cells through an amiloride-blockable sodium channel. Taste bud cells utilizing this mechanism exclusively activate N fibers, which are involved in the control of NaCl intake. A different mechanism for the detection of NaCl and other electrolytes is utilized by taste bud cells that activate H fibers.

Persistence of taste buds in denervated fungiform papillae

Taste buds in hamster fungiform papillae persist in an atrophic state for as long as 330 days after chorda tympani denervation or 50 days after combined chorda tympani-lingual nerve resection. Although taste bud structure depends on innervation, there is no absolute neural requirement for taste bud survival.

Taste-responsive neurons and their locations in the solitary nucleus of the hamster

The solitary nucleus (nucleus tractus solitarii), the first central relay for taste in mammals, was studied anatomically and physiologically in the golden hamster (Mesocricetus auratus). Activity of neurons to anterior tongue stimulation with sucrose, NaCl and KCl were extracellularly recorded. Electrolytic lesions or horseradish peroxidase deposits allowed subsequent localization of recording sites. Anterior tongue taste-responsive sites were restricted to a very small part of the rostral pole of the solitary nucleus, which is about 3% of the entire nucleus. Sites were confined to the rostral-central and rostral-lateral subdivisions of Whitehead, which contain a number of morphological cell types. Some chemotopic organization was seen with multi-unit recordings, with NaCl-selective sites concentrated rostrally and sucrose- and KCl-selective sites concentrated caudally. Sites with broad sensitivity were distributed throughout the gustatory region. Single neural units showing inhibition to taste stimuli, units highly reactive to all three stimuli, and units with high spontaneous rates were seen in the solitary nucleus, as well as units that responded very selectively and had low spontaneous rates. Single units with similar response profiles to sucrose, NaCl and KCl were not segregated to separate restricted locations within the taste-reactive region; their distributions overlapped. In the hamster, neurons in the anterior tongue taste region of the solitary nucleus process taste quality information in diverse ways. Highly reactive non-specific neurons, neurons that show inhibition, and neurons with high spontaneous rates are more frequently observed in the solitary nucleus than in the afferent input fibers of the chorda tympani nerve. The small region of the rostral pole enclosing taste-responsive neurons is complexly organized in relation to taste quality and contains a number of morphological cell types whose functional role in taste is not yet known.

The distinctiveness of ionic and nonionic bitter stimuli.

The diverse chemical structures of stimuli that are bitter to humans suggest a need for multiple bitter receptors. Reactions of golden hamsters (Mesocricetus auratus) to 1 mM quinine hydrochloride, 3 mM denatonium benzoate, 180 mM magnesium sulfate, 30-100 mM caffeine, and 1-1.5 mM sucrose octaacetate (SOA) were studied to address whether there are multiple sensations elicited by bitter stimuli. Methods included behavioral generalization of LiCl-induced conditioned taste aversions (CTAs), intake preference tests, and electrophysiological recordings from the chorda tympani (CT) nerve. The five compounds, all bitter to humans, were all innately aversive to hamsters. CTA for the ionic quinine.HCl, denatonium benzoate, and MgSO(4) mutually cross-generalized and these ionic compounds were effective CT stimuli. Yet, the hamsters were much less sensitive to denatonium than humans, requiring a 100,000 times higher concentration for detection. CTA for nonionic caffeine and SOA did not cross-generalize to quinine or the other two ionic stimuli and these nonionic compounds were not effective CT stimuli. SOA and caffeine may elicit aversive reflexes or systemic reactions rather than taste sensations in the animals. Thus, the three ionic and two nonionic compounds form separate aversive stimulus classes in hamsters, neither of which appears to be a close homologue of the human bitter taste.

Taste responses to mixtures: analytic processing of quality.

The tastes of 100 mM sodium chloride (NaCl), 100 mM sucrose, and 1 mM quinine hydrochloride in mixtures were investigated in golden hamsters (Mesocricetus auratus) with a conditioned taste aversion (CTA) paradigm. CTAs, established in golden hamsters by injection of lithium chloride, were quantified as percent suppression of control 1-hr stimulus intake. CTAs for 10 of 15 stimulus pairs with common components symmetrically cross-generalized, suggesting that component qualities were recognized in binary and ternary mixtures. However, CTAs to quinine were hardly learned and were weakly expressed when quinine was mixed with NaCl, and generalizations from multiple to single stimuli were stronger than vice versa (i.e., asymmetric). The behaviors reflect peripheral inhibition and/or central mixture suppression. Nonetheless, components retain their distinct qualities in mixtures, suggesting that taste processing is analytic.

Opponent effects of quinine and sucrose on single fiber taste responses of the chorda tympani nerve.

Responses of single chorda tympani fibers to mixtures of taste stimuli were studied in the golden hamster (Mesocricetus auratus). Sucrose-best neurons showed significant suppression to quinine-sucrose mixtures compared to sucrose alone. Quinine may exert its effect as an opponent stimulus in the receptor cells at the second messenger level. This suppression may make bitter quinine more readily detected when embedded in mixtures with sweeteners.

Information processing in mammalian gustatory systems

The taste system has multiple functions that are carried by three cranial nerves. It is now apparent that these functions cannot be accommodated by a single coding mechanism for taste quality. A current view emphasizes the likely existence of coding channels activated by specific sets of receptors.

Peripheral gustatory processing of sweet stimuli by golden hamsters.

Behaviors and taste-nerve responses to bitter stimuli are linked to compounds that bind T2 receptors expressed in one subset of taste-bud receptor cells (TRCs); and behavioral and neural responses to sweet stimuli are linked to chemical compounds that bind a T1 receptor expressed in a different TRC subset. Neural and behavioral responses to bitter-sweet mixtures, however, complicate the ostensible bitter and sweet labeled lines. In the golden hamster, Mesocricetus auratus, quinine hydrochloride, the bitter prototype, suppresses chorda tympani (CT) nerve responses to the sweet prototype: sucrose. This bitter-sweet inhibition was tested with concentration series of sucrose and dulcin, a hydrophobic synthetic sweetener that hamsters behaviorally cross-generalize with sucrose. Dulcin, sucrose and other sweeteners activate one subset of CT fibers: S neurons; whereas, quinine activates a separate subset of CT fibers: E neurons. Whole-nerve and S-neuron CT responses to a sweetener concentration series, mixed with 0, 1, 3 and 10 mM quinine, were measured for 0-2.5 s transient and/or 2.6-10 s steady-state response periods. Ten-sec total single-fiber records, aligned at response onset, were averaged for 100 ms bins to identify response oscillations. Quinine inhibition of dulcin and sucrose responses was identical. Each log molar increment in quinine resulted in equivalent declines in response to either sweetener. Furthermore, sucrose response decrements paralleled response increments in quinine-sensitive CT neurons to the same quinine increases. A 1.43 Hz bursting rhythm to the sweeteners was unchanged by quinine inhibition or decreases in sweetener concentration. Taste-bud processing, possibly between-cell inhibition and within-cell negative feedback, must modify signals initiated by T1 receptors before they are transmitted to the brain.

Effects of chlorhexidine on human taste perception

Chlorhexidine, a bis-cationic biguanide antiseptic, greatly reduces the perceived intensity of the salty prototype sodium chloride and may prove to be an important probe of mechanisms that underlie the human salty taste quality. Chlorhexidine, which tastes bitter, also reduces quinine hydrochloride taste intensity, but neither sweet sucrose nor sour citric acid is affected. Perceptual intensity rating and quality identification were measured for human subjects before and for 30 min following treatment with 1.34 mM chlorhexidine gluconate. In one experiment, test stimuli were the taste-quality prototypes; in a second experiment, stimuli were series of sodium, halide and sulfate salts. Experiment 1 showed a single 3-min chlorhexidine treatment resulted in reductions in taste intensity that persisted for at least 30 min. Experiment 2 showed a single 2-min chlorhexidine treatment reduced perceptual intensities of halide and sulfate salts except those with divalent cations. Chlorhexidine impaired identification of the salty quality and produced a bitter quality in nonbitter salts and impaired identification of the bitter quality of quinine, but not bitter salts. The specific effect of chlorhexidine on the bitterness of quinine suggests it may bind to the same receptor as quinine. The ability of chlorhexidine to specifically disrupt saltiness of a wide range of salts is consistent with proposed peripheral transduction mechanisms for the salty quality that involve transepithelial ion transport.

study of taste perception

Taste stimulus identification was studied in order to more thoroughly examine human taste perception. Ten replicates of an array of 10 taste stimuli—NaCl, KCl, Na glutamate, quinine.HCl, citric acid, sucrose, aspartame, and NaCl-sucrose, acid-sucrose, and quinine-sucrose mixtures—were presented to normal subjects for identification from a list of corresponding stimulus names. Because perceptually similar substances are confused in identification tasks, the result was ataste confusion matrix. Consistency of identification for the 10 stimuli (T10) and for each stimulus pair (T2) was quantified with measures derived from information theory. Forty-two untrained subjects made an average of 57.4% correct identifications. An averageT10 of 2.25 of the maximum 3.32 bits and an averageT2 of 0.84 of a maximum 1.0 bit of information were transmitted. In a second experiment, 40 trained subjects per-formed better than 20 untrained subjects. The results suggested that the identification procedure may best be used to assess taste function following 1–2 training replicates. The patterns of taste confusion indicate that the 10 stimuli resemble one another to varying extents, yet each can be considered perceptually unique.