Amiloride-insensitive units of the chorda tympani nerve are necessary for normal ammonium chloride detectability in the rat

In spite of its common use as a standard stimulus in peripheral nerve recordings, relatively little is known about the psychophysics of NH-sub-4Cl taste. Rats' detection threshold for this salt was tested under a variety of conditions, including amiloride (100 muM) treatment and bilateral chorda tympani (CT) nerve transection. Detectability was measured with a 2-lever operant discrimination procedure used previously to measure detection thresholds for NaCl and KCl. Although NH-sub-4Cl and KCl appear to share a common taste quality and transduction mechanism, the logistic function and threshold for NH-sub-4Cl were found to be more similar to those of NaCl than to those of KCl. Like that of KCl, however, the detection threshold for NH4Cl increased significantly with CT transection (0.54 log-sub-1-sub-0 units, p < .004), but not with amiloride adulteration. This finding supports the hypothesis that the CT is necessary for normal salt detection regardless of stimulus, and suggests that amiloride does not appreciably impact responses to nonsodium salts at the behavioral level.     

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.     

Anion size does not compromise sodium recognition by rats after acute sodium depletion

Amiloride-insensitive sodium taste transduction is severely limited by large anions (i.e., gluconate). We found that in a brief-access taste test, sodium-depleted rats exhibited similar levels of increased licking to several sodium salts regardless of anion but did not increase licking to nonsodium salts compared with water. The enhanced licking of sodium salts was abolished in the presence of amiloride. These results suggest that the amiloride-sensitive taste transduction pathway is not only necessary but that it is also sufficient for sodium identification in rats. Sodium-depleted rats tested with amiloride initiated significantly more trials than nondepleted rats; hence, appetitive behavior was mildly potentiated by depletion, even in the absence of a sodium taste cue. Overall, these findings provide compelling support for the primacy of the amiloride-sensitive taste transduction mechanism and its associated neural pathway in the recognition of the sodium cation.     

Perceptual characteristics of the amiloride-suppressed sodium chloride taste response in the rat

The contribution of amiloride-sensitive membrane components to the perception of NaCl taste was assessed by using a conditioned taste aversion procedure. Eight independent groups of adult rats were conditioned to avoid either 0.1M NaCl, 0.5M NaCl; 0.1M NH4Cl, or 1.0M sucrose while their tongues were exposed either to water or to the sodium transport blocker amiloride hydrochloride. In contrast to rats exposed to water during conditioning, rats exposed to amiloride were unable to acquire a conditioned taste aversion to 0.1M NaCl. Differences in the acquisition of taste aversions between the amiloride- and nonamiloride-treated groups were not apparent when the conditioned stimulus (CS) was 0.5M NaCl, 0.1M NH4Cl, or 1.0M sucrose. Although the magnitude of the 0.5M NaCl aversion was similar between amiloride- and non-amiloride-treated rats, the perceptual characteristics of the CS differed between groups. Analyses of stimulus generalization gradients revealed that amiloride-treated rats generally avoided all monochloride salts after conditioning to 0.5M NaCl but not nonsodium salts or nonsalt stimuli. In contrast, rats not treated with amiloride only generalized the 0.5M NaCl aversion to sodium salts. No differences in generalization gradients occurred between groups when the CS was 0.1M NH4Cl or 1.0M sucrose. These findings suggest that the "salty" taste of NaCl is primarily related to the amiloride-sensitive portion of the functional taste response in rats. Conversely, the portion of the NaCl response insensitive to amiloride appears to have "sour-salty" perceptual characteristics and does not appear to be perceived as being salty.     

Psychophysical investigations of cetylpyridinium chloride in rats: its inherent taste and modifying effects on salt taste

Salts are transduced by at least 2 mechanisms: (a) antagonized by amiloride and (b) antagonized by cetylpyridinium chloride (CPC). The authors report on 4 behavioral experiments in rats that characterize the orosensory properties of CPC itself as well as its effect in suppressing the intensity of NaCl and KCl taste. Experiments 1 and 2 indicated that CPC has a quinine-like taste quality. Experiments 3 and 4 demonstrated that the recognition of KCl, but not NaCl, is modestly reduced by mixture with CPC. However, control experiments call into question the mechanism of the salt suppression of CPC, because both CPC-salt and quinine-salt mixtures had similar effects. The relevance of these studies for understanding salt and bitter taste coding is discussed.     

Lack of amiloride sensitivity in SHR and WKY glossopharyngeal taste responses to NaCl.

To explore possible functional strain differences in taste receptors located on the posterior tongue, we recorded electrophysiological taste responses from the glossopharyngeal nerve of spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats. Multifiber responses to a concentration series (0.5 M to 2.0 M) of NaCl, KCl and NH4Cl were recorded before and after lingual application of the epithelial sodium transport blocker, amiloride. Responses to a concentration series (0.0025 M to 0.1 M) of quinine hydrochloride were also recorded. When expressed relative to the 0.5-M NH4Cl response, responses to the monochloride salts were equivalent between SHR and WKY. Surprisingly, NaCl responses were not suppressed by the sodium transport blocker, amiloride. This is in direct contrast to the dramatic suppression observed in the chorda tympani. Also, relative responses to quinine were greater in the glossopharyngeal nerve of SHR than WKY. These results indicate that taste receptors innervated by the glossopharyngeal nerve lack amiloride sensitivity and that posterior taste receptor function to monochloride salts is equivalent between SHR and WKY.     

Dietary sodium deprivation reduces gustatory neural responses of the parabrachial nucleus in rats

Acute sodium depletion induced by furosemide reduces gustatory responses of parabrachial nucleus (PBN) neurons to 0.3-0.5M NaCl in rats. However, in the rat nucleus of the solitary tract (NST), where taste-responsive cells project to the PBN, acute sodium depletion and dietary sodium deprivation elicit different response profiles to lingual NaCl stimulation. To examine the effect of dietary sodium deprivation on the responses of PBN gustatory neurons, we observed the taste responses of the PBN neurons to the four taste qualities and serial concentrations of NaCl in 15-day dietary sodium-deprived and control rats. The results showed that sodium deprivation reduced the responses of PBN taste neurons to 0.1-1.0M NaCl, but not to other tastants. Based on the analyses classified by best-stimulus categories, the number of NaCl-best neurons decreased from 68% to 45% following dietary sodium deprivation, and the responses of the NaCl-best neurons to 0.03-1.0M NaCl were significantly inhibited. Multidimensional scaling illustrated that sodium deprivation increased the similarity of the response profiles of the NaCl-best neurons. These findings suggest that dietary sodium deprivation might modulate sodium intake via increasing aversive threshold for salt rather enhancing salt discrimination.     

Alterations of salt taste perception in the developing rat

Behavioral correlates of changing neurophysiological taste sensitivities during development were assessed with a conditioned taste aversion procedure. Young rats (age 25-30 days) avoided 0.1M monochloride salts and 1.0M sucrose reliably less than adults (age 90-105 days), but the two groups did not differ when the conditioned stimulus (CS) was 0.1M citric acid. Analyses of generalization gradients revealed that young rats were unable to discriminate among the tastes of NaCl, NH4Cl, and KCl, whereas adults readily made such discriminations. Both age groups had similar generalization gradients when the CS was 1.0M sucrose or 0.1M citric acid. These data indicate that quantitative and qualitative aspects of salt taste perception alter with age. Furthermore, the behavioral changes noted in the present study correspond closely with previous findings from developmental studies of neurophysiological taste responses.     

Artificial neural networks estimate the contribution of taste neurons to coding

Taste qualities are believed to be coded in the activity of populations of taste neurons. However, it is not clear whether all neurons are equally responsible for coding. To clarify the point the relative contribution of each taste neuron to coding was assessed by constructing simple three-layer neural networks with input neurons that represent cortical taste neurons of the rat. The networks were trained by the back-propagation learning algorithm to classify the neural response patterns to the basic taste stimuli (sucrose, HCl, quinine-hydrochloride, and NaCl). The networks had four output neurons representing the basic taste qualities, the values of which provide a measure for similarity of test stimuli to the basic taste stimuli. We estimated relative contributions of input neurons to the taste discrimination of the network by examining their significance S(j), which is defined as the sum of the absolute values of the connection weights from the jth input neuron to the hidden layer. When the input neurons with a smaller S(j) (e.g., 15 out of 39 input units) were "pruned" from the trained network, the ability of the network to discriminate the basic taste qualities was not greatly affected. On the other hand, the taste discrimination of the network progressively deteriorated much more rapidly with pruning of input neurons with a larger S(j). These results suggest that cortical taste neurons differentially contribute to the coding of taste qualities. Input neurons with a larger S(j) tended to be with a larger variation of neural discharge rates to the basic taste stimuli. The variation of neural discharges may be important in the coding of taste qualities.     

Taste-responsive neurons of the glossopharyngeal nerve of the rat

1. Taste sensibilities of neurons in mammalian glossopharyngeal nerves have been inadequately studied, although they innervate the majority of taste buds and may provide unique taste information. 2. Extracellular responses of glossopharyngeal neural units to taste stimuli infused into foliate or vallate papillae were recorded in anesthetized rats. A 0.3-ml/min infusion of stimuli into papillae resulted in short-latency, 5-s nerve-impulse rates that approached 10 times the response rates observed using less invasive means of stimulation. 3. Sucrose, Na saccharin, NaCl, NH4Cl, KCl, HCl, citric acid, acetic acid, MgSO4, and quinine.HCl were effective stimuli for glossopharyngeal neurons at concentrations that have behavioral significance. 4. Response spectra for individual neural units with either foliate or vallate receptive fields fell into three clusters. Forty-six percent were A units that responded most strongly to acids and chloride salts, NH4Cl being the most effective; neither quinine nor sucrose was effective. Twenty-three percent were S units that responded to sugars and saccharin; quinine, salts, and acids were not effective. Thirty-one percent were Q units that responded to quinine; neither NaCl, HCl, nor sucrose was effective stimulus for these fragile units. 5. Glossopharyngeal A neural units were more sensitive to 1 mM HCl than were electrolyte-sensitive H units of the chorda tympani, although both respond generally to salts and acids. Units relatively specific for sodium salts (N units), which are common in the chorda tympani nerve, were not found in the glossopharyngeal nerve, which explains losses in sodium-specific behavior after cutting only the chorda tympani nerve. 6. Q units were the only glossopharyngeal neural units that responded significantly to quinine, and units with similar response spectra do not occur in the chorda tympani nerve. Q units probably mediate aversive reflexes to quinine that are eliminated by cutting only the glossopharyngeal nerve. Glossopharyngeal S neural units were more sensitive to sucrose and are more common than their counterparts in the chorda tympani, although it is not known how they might compare with sugar-sensitive units in the greater superficial petrosal nerve. 7. These data strongly suggest that posterior taste bud fields innervated by the glossopharyngeal nerve are specialized for functions different from those of anterior taste bud fields innervated by the facial nerve.     

Sodium deficient rats are unmotivated by sodium chloride solutions mixed with the sodium channel blocker amiloride.

Rats were made sodium deficient by furosemide injection and then offered 20 min of access to 0.05 M NaCl mixed with the sodium channel blocker amiloride. Compared with a sodium deficient control group that was also offered 0.05 M NaCl, these rats drank very little. A subsequent test conducted in the same manner with 20 min of access to 0.3 M NaCl mixed with amiloride produced similar results. It is concluded that amiloride blocks the neural information required for generating the attractive taste of NaCl to the sodium deficient rat.     

Development of taste responses in rat nucleus of solitary tract

Extracellular responses from neurons in the nucleus of the solitary tract (NST) were studied in rats aged 5 days to adulthood during chemical stimulation of the tongue with monochloride salts, citric and hydrochloric acids, sucrose, sodium saccharin, and quinine hydrochloride. Multiunit taste responses were recorded in rats at 5-7 days of age and single-unit responses were recorded from 111 neurons in four other age groups of 14-20 days, 25-35 days, 50-60 days, and adult. NST neurons in rats aged 5-7 days consistently responded to relatively high concentrations (0.5 M) of NH4Cl and KCl and to citric and hydrochloric acid. However, they often did not respond to 0.5 M NaCl or to 0.1 M NH4Cl. Single NST neurons in rats aged 14 days and older characteristically responded to all 0.1 and 0.5 M salts and to both acids. At least 75% of neurons also responded to sucrose and sodium saccharin, and 46% responded to all of these stimuli and quinine hydrochloride. After 14 days, no developmental changes occurred in the number of stimuli to which neurons responded. There were substantial developmental alterations in the response magnitudes to some chemical stimuli. Average response frequencies increased after 35 days of age for 0.1 and 0.5 M NaCl, LiCl, KCl, and for sucrose and sodium saccharin. Response frequencies for NH4Cl, citric and hydrochloric acid, and quinine hydrochloride, however, did not change throughout development. The proportion of single NST neurons that responded maximally to specific monochloride salts did not change during development. Most single neurons in all age groups responded equally well to NH4Cl, NaCl, and LiCl. No NST neuron responded maximally to KCl. There were also no developmental differences in response latencies in rats aged 14 days and older. Response frequencies of second-order NST neurons generally reflect changes in responses from the primary afferent, chorda tympani fibers, throughout development; however, the increases in salt response frequencies from NST neurons occur comparatively later in development. Furthermore, at all ages, the taste responses to monochloride salts include higher response frequencies and a general loss in response specificity in NST compared to chorda tympani neurons.(ABSTRACT TRUNCATED AT 400 WORDS)     

Behavioral and neural gustatory responses in rabbit

To provide more information on a potentially valuable preparation for studies in taste and appetite, we have examined the taste preferences (and aversions) and chorda tympani sensitivity of the rabbit. Adult male New Zealand rabbits were given a two-bottle preference test between water and various molar concentrations of NaCl, KCl, sucrose, sodium saccharin, quinine hydrochloride and HCl. The rabbits exhibited the expected preferences for sucrose and aversions for quinine and HCl. Unexpectedly, however, the rabbits exhibited only a mild preference for NaCl, a stronger preference for KCl, and an aversion to sodium saccharin. Multiunit discharges of the chorda tympani nerve to the same taste stimuli indicated that the anterior tongue receptors are acutely sensitive to KCl, NaCl and quinine, but not to sucrose, HCl and saccharin. The chorda tympani was more responsive to KCl than to NaCl. Dilute concentrations of both NaCl and sodium saccharin elicited a two-component response consisting of an immediate excitatory phase followed by a tonic inhibitory phase. This complex response pattern of the whole nerve to NaCl and sodium saccharin is discussed in relation to the impulse frequencies in hypothesized water-sensitive and salt-sensitive fibers. Both the behavioral and neural data are discussed in relation to similar data obtained in rat and hamster.     

Amiloride sensitivity of the chorda tympani response to sodium chloride in sodium-depleted Wistar rats.

Peripheral gustatory mechanisms that may contribute to the expression of sodium (Na) appetite have been a focus of interest for many years. Because amiloride-sensitive Na transport is involved in the generation of neural signals in response to NaCl stimulation, the present study assessed whether changes in amiloride sensitivity of the neural response to NaCl accompany the induction of a Na appetite in the rat. Na deprivation was achieved by acute depletion with the diuretic furosemide. The magnitude of the whole-nerve chorda tympani response to 0.5 M NaCl was reduced in Na-depleted, compared with Na-replete, rats, which provides qualified support for previous reports that the induction of a Na appetite is associated with reduced neural responses to NaCl. However, changes in sensitivity to the specific Na channel blocker amiloride hydrochloride as a result of Na depletion were not evident. These findings suggest that the behavioral and neural changes that occur after Na depletion are not based on changes in amiloride sensitivity in the taste bud.     

Chemosensory function of amphibian skin: integrating epithelial transport, capillary blood flow and behaviour

Terrestrial anuran amphibians absorb water across specialized regions of skin on the posterioventral region of their bodies. Rapid water absorption is mediated by the insertion of aquaporins into the apical membrane of the outermost cell layer. Water moves out of the epithelium via aquaglyceroporins in the basolateral membrane and into the circulation in conjunction with increased capillary blood flow to the skin and aquaporins in the capillary endothelial cells. These physiological responses are activated by intrinsic stimuli relating to the animals' hydration status and extrinsic stimuli relating to the detection of osmotically available water. The integration of these processes has been studied using behavioural observations in conjunction with neurophysiological recordings and studies of epithelial transport. These studies have identified plasma volume and urinary bladder stores as intrinsic stimuli that activate the formation of angiotensin II (AII) to stimulate water absorption behaviour. The coordinated increase in water permeability and capillary blood flow appears to be mediated primarily by sympathetic stimulation of beta adrenergic receptors, although the neurohypopyseal hormone arginine vasotocin (AVT) may also play a role. Extrinsic stimuli relate primarily to the ionic and osmotic properties of hydration sources. Toads avoid NaCl solutions that have been shown to be harmful in acute exposure, approx. 200-250 mm. The avoidance is partially attenuated by amiloride raising the hypothesis that the mechanism for salt detection by toads resembles that for salt taste in mammals that take in water by mouth. In this model, depolarization of the basolateral membrane of taste cells is coupled to afferent neural stimulation. In toad skin we have identified innervation of skin epithelial cells by branches of spinal nerves and measured neural responses to NaCl solutions that elicit behavioural avoidance. These same concentrations produce depolarization of the basolateral membrane in isolated epithelial preparations. As with salt taste in mammals, the neural responses and depolarization of basolateral membrane potential are partially inhibited by amiloride. In addition, toads are more tolerant of sodium gluconate solution which is consistent with the phenomenon in mammalian taste physiology termed the anion paradox in which sodium salts with larger molecular weight anions produce a reduced intensity of salt taste. Finally, toads also avoid concentrated solutions of a non-electrolyte, mannitol, which differs from NaCl solutions in not affecting transepithelial conductance and requires a longer time to depolarize the basolateral membrane. Osmotic stimuli may mediate sensory processes for longer term detection of conditions with low water potential while ionic stimuli are more important for shorter term analysis of rehydration sources.     

Neural representation of bitter taste in the nucleus of the solitary tract

Based on the molecular findings that many bitter taste receptors (T2Rs) are expressed within the same receptor cells, it has been proposed that bitter taste is encoded by the activation of discrete neural elements. Here we examined how a variety of bitter stimuli are represented by neural activity in central gustatory neurons. Taste responses (spikes/s) evoked by bathing the tongue and palate with intensity-matched concentrations (in M) of 2 sugars (0.32 sucrose and 0.5 D-fructose), ethanol (40%), 4 salts (0.01 NaCl, 0.008 NaNO(3), 0.01 MgCl(2), and 0.05 KCl), 2 acids (0.003 HCl and 0.005 citric acid), and 10 bitter ligands (0.007 quinine-HCl, 0.015 denatonium benzoate, 0.003 l-cysteine, 0.001 nicotine, 0.005 strychnine-HCl, 0.04 tetraethylammonium chloride, 0.03 atropine-SO(4), 0.005 brucine-SO(4), 0.03 papaverine-HCl, and 0.009 sparteine) were recorded from 51 neurons in the nucleus of the solitary tract of anesthetized rats. Cluster analysis was used to categorize neurons into types based on responses to sucrose, NaCl, HCl, and quinine-HCl. Three groupings emerged: type S (responded optimally to sweets), type N (sodium-optimal), and type H/Q (responded robustly to bitters, acids, and salts). Multivariate analyses revealed that across-neuron patterns of response among bitter stimuli were strongly correlated. However, neural type H/Q, which was most responsive to bitter tastants, was not differentially sensitive to bitter stimuli and Na(+) salts, which rats perceive as distinct. Thus central neurons most responsive to bitter substances receive significant input from receptors that mediate other tastes, indicating that bitter stimuli are not represented by activity in specifically tuned neurons.     

Gustatory neuron types in hamster brain stem

In general, mammalian taste neurons are broadly responsive to stimuli representing different taste qualities. In the hamster, this breadth of tuning increases systematically from peripheral to successively higher brain stem neurons. Some investigators have classified taste-responsive neurons into "best-stimulus" categories on the basis of which of the four basic stimuli (sucrose, NaCl, HCl, or quinine hydrochloride) elicits the maximum response. However, attempts by others to demonstrate the existence of taste neuron types in the chorda tympani nerve and medulla of the rat using hierarchical cluster analysis have not been successful, resulting in the conclusion that there are no neuron types in the rat gustatory system. The present study was designed to look at the question of neuron types in the hamster, a species with a broader range of gustatory sensitivities to anterior tongue stimulation. Responses of 30 neurons in the nucleus tractus solitarius (NTS) and 31 neurons in the parabrachial nuclei (PbN) of the hamster to an array of 18 stimulus compounds were recorded extracellularly. The similarities of the neural response profiles of these cells at each synaptic level were compared using multivariate statistical techniques. The possiblee grouping of cells on the basis of similarities in their response functions was examined with hierarchical cluster analysis, and the relationships among these response functions were examined with multidimensional scaling. The results of the cluster analysis suggested that at both the NTS and PbN, there are three clusters of neural response profiles. These three clusters of response profiles are characterized at both synaptic levels by their predominant sensitivity to 1) sucrose and other sweet-tasting compounds, 2) sodium salts, and 3) nonsodium salts and acids. Representation of these neurons in a two-dimensional space yielded three nonoverlapping groups of cells in both the NTS and PbN, corresponding to the three groups identified by the hierarchical cluster solution. Classification of taste neurons either by their best stimulus or by other criteria has been criticized on the grounds that it may constitute an arbitrary division of a continuous population of neurons. The techniques of numerical taxonomy, which take the cells' variability into account, also result in a grouping of taste cells into classes. These taxonomic classes agree in most instances (80% in NTS and 80.6% in PbN) to a best-stimulus classification. The failure of some investigators to find types of neural response profiles in the rat gustatory system may be the result of species differences in taste sensitivity as well as differences in the statistical procedures employed.     

The effect of amiloride on operantly conditioned performance in an NaCl taste detection task and NaCl preference in C57BL/6J mice

A 2-response operant taste discrimination procedure, modified to assess taste sensitivity in water-restricted C57BL/6J mice, revealed a detection threshold of 0.065 M sodium chloride. Amiloride increased the threshold by approximately 1 log10 unit. These results are the first to demonstrate the necessity of the amiloride-sensitive taste transduction pathway in the normal detection of low concentrations of sodium chloride in mice and provide a functional context in which to evaluate electrophysiological findings. Two-bottle preference tests performed with these mice and additional naive mice revealed only marginal, if any, effects of amiloride on salt intake behavior, highlighting the importance of considering the relative attributes and limitations of different behavioral assays of taste function.     

Gustatory neuron types in rat geniculate ganglion

We used extracellular single-cell recording procedures to characterize the chemical and thermal sensitivity of the rat geniculate ganglion to lingual stimulation, and to examine the effects of specific ion transport antagonists on salt transduction mechanisms. Hierarchical cluster analysis of the responses from 73 single neurons to 3 salts (0.075 and 0.3 M NaCl, KCl, and NH(4) Cl), 0.5 M sucrose, 0.01 M HCl, and 0.02 M quinine HCl (QHCl) indicated 3 main groups that responded best to either sucrose, HCl, or NaCl. Eight narrowly tuned neurons were deemed sucrose-specialists and 33 broadly tuned neurons as HCl-generalists. The NaCl group contained three identifiable subclusters: 18 NaCl-specialists, 11 NaCl-generalists, and 3 QHCl-generalists. Sucrose- and NaCl-specialists responded specifically to sucrose and NaCl, respectively. All generalist neurons responded to salt, acid, and alkaloid stimuli to varying degree and order depending on neuron type. Response order was NaCl > HCl = QHCl > sucrose in NaCl-generalists, HCl > NaCl > QHCl > sucrose in HCl-generalists, and QHCl = NaCl = HCl > sucrose in QHCl-generalists. NaCl-specialists responded robustly to low and high NaCl concentrations, but weakly, if at all, to high KCl and NH(4) Cl concentrations after prolonged stimulation. HCl-generalist neurons responded to all three salts, but at twice the rate to NH(4) Cl than to NaCl and KCl. NaCl- and QHCl-generalists responded equally to the three salts. Amiloride and 5-(N,N-dimethyl)-amiloride (DMA), antagonists of Na(+) channels and Na(+)/H(+) exchangers, respectively, inhibited the responses to 0.075 M NaCl only in NaCl-specialist neurons. The K(+) channel antagonist, 4-aminopyridine (4-AP), was without a suppressive effect on salt responses, but, when applied alone in solution, it evoked a response in many HCl-generalists and one QHCl-generalist neuron so tested. Of the 39 neurons tested for their sensitivity to temperature, 23 responded to cooling and chemical stimulation, and 20 of these neurons were HCl-generalists. Moreover, the responses to the four standard stimuli were reduced progressively at lower temperatures in HCl- and QHCl-generalist neurons, but not in NaCl-specialists. Thus sodium channels and Na(+)/H(+) exchangers appear to be expressed exclusively on the membranes of receptor cells that synapse with NaCl-specialist neurons. In addition, cooling sensitivity and taste-temperature interactions appear to be prominent features of broadly tuned neuron groups, particularly HCl-generalists. Taken all together, it appears that lingual taste cells make specific connections with afferent fibers that allow gustatory stimuli to be parceled into different input pathways. In general, these neurons are organized physiologically into specialist and generalist types. The sucrose- and NaCl-specialists alone can provide sufficient information to distinguish sucrose and NaCl from other stimuli, respectively.