Steven J. St. John

Behavioral Discrimination between Quinine and KCl Is Dependent on Input from the Seventh Cranial Nerve: Implications for the Functional Roles of the Gustatory Nerves in Rats

The rat glossopharyngeal nerve (GL), which innervates posterior tongue taste buds, contains several physiologically defined taste fiber types; at least one type is primarily responsive to certain alkaloids (such as quinine), and another is primarily responsive to acids and salts. In contrast, the chorda tympani (CT), which innervates anterior tongue taste buds, does not appear to contain fibers that differentially respond to quinine relative to salts and acids. It was therefore predicted that GL transection should disrupt behavioral discriminations between quinine and either acids or salts. Water-restricted rats were trained to press one of two levers if a sampled taste stimulus was quinine (0.1–1.0 mm) and the second lever if the sampled stimulus was KCl (0.1–1.0 m). Sham surgery, GL transection, and sublingual and submaxillary salivary gland extirpation were found to have no effect relative to presurgical performance. Both CT transection and combined GL and CT transection caused a substantial and approximately equal decrement in discrimination performance. Removal of the gustatory branches of the seventh cranial nerve [CT and greater superficial petrosal (GSP)] nearly eliminated the discrimination of the taste stimuli, and combined transection of the CT, GL, and GSP unequivocally reduced performance to chance levels. Although these findings were not presaged by the known electrophysiology, they nonetheless compare favorably with other studies reporting little effect of GL transection on behavioral responses to quinine. These results, in the context of other discrimination studies reported in the literature, suggest that, in rats, the neural coding of taste quality depends primarily on the input of the facial nerve.

Combined, but not single, Gustatory nerve transection substantially alters taste-guided licking behavior to quinine in rats

On the basis of electrophysiological studies, the glossopharyngeal nerve (GL) is far more responsive to quinine than the chorda tympani (CT) or greater superficial petrosal (GSP) nerves. The licking behavior of water-deprived rats to quinine (0.03-3.0 mM) and distilled water (10-s trails) was tested before and after various nerve transections. GL+CT section caused a substantial reduction in responsiveness. GSP+CT section had a moderate effect, and GL section alone produced only marginal impairments. Control, partially desalivated, and CT-sectioned rats were unaffected. Thus, the GL is not necessary for normal unconditioned taste-guided appetitive responsiveness to quinine, but the collective input from the GSP and CT is necessary and most likely sufficient. These data suggest that the quinine-evoked input of the GL and CT converge centrally.

Role of taste in the microstructure of quinine ingestion by rats

The microstructure of the licking behavior of water-deprived rats presented with either water or quinine during 45-min single-bottle tests was analyzed. The chorda tympani (CT) and glossopharyngeal (GL) nerves, which innervate the taste buds of the tongue, were transected in deeply anesthetized rats to discern their contribution to the behavioral pattern of quinine drinking. Rats were presurgically habituated to the testing protocol and postsurgically tested first with water and then novel 0.2 mM quinine-HCl in a subsequent session. The substantial decrease in intake observed in sham-operated controls (n = 16) when quinine was the stimulus was entirely a function of a decrease in lick volume and burst size (a run of licks with interlick intervals <1 s). Contrary to the intake-suppressing effects of quinine, pause duration decreased and burst number increased. Combined transection of the CT and GL (n = 6) strikingly opposed all of these quinine-induced behavioral changes, whereas CT transection (n = 7) was without effect and GL transection (n = 8) had an intermediate influence. These results suggest that taste acts more on neural circuits governing burst termination as opposed to burst initiation, which, in turn, appears to be more sensitive to signals related to physiological state. These findings are discussed in terms of other known nerve transection effects on quinine responsiveness, and the implications of the microstructural results are considered with respect to probabilistic as opposed to deterministic control of licking behavior.The microstructure of the licking behavior of water-deprived rats presented with either water or quinine during 45-min single-bottle tests was analyzed. The chorda tympani (CT) and glossopharyngeal (GL) nerves, which innervate the taste buds of the tongue, were transected in deeply anesthetized rats to discern their contribution to the behavioral pattern of quinine drinking. Rats were presurgically habituated to the testing protocol and postsurgically tested first with water and then novel 0.2 mM quinine-HCl in a subsequent session. The substantial decrease in intake observed in sham-operated controls ( n = 16) when quinine was the stimulus was entirely a function of a decrease in lick volume and burst size (a run of licks with interlick intervals <1 s). Contrary to the intake-suppressing effects of quinine, pause duration decreased and burst number increased. Combined transection of the CT and GL ( n = 6) strikingly opposed all of these quinine-induced behavioral changes, whereas CT transection ( n = 7) was without effect and GL transection ( n = 8) had an intermediate influence. These results suggest that taste acts more on neural circuits governing burst termination as opposed to burst initiation, which, in turn, appears to be more sensitive to signals related to physiological state. These findings are discussed in terms of other known nerve transection effects on quinine responsiveness, and the implications of the microstructural results are considered with respect to probabilistic as opposed to deterministic control of licking behavior.

Temporal and qualitative dynamics of conditioned taste aversion processing : Combined generalization testing and licking microstructure analysis

The pattern of licking microstructure during various phases of a conditioned taste aversion (CTA) was evaluated. In Experiment 1, rats ingested lithium chloride (LiCl) for 3 trials and were then offered sodium chloride (NaCl) or sucrose on 3 trials. A CTA to LiCl developed and generalized to NaCl but not to sucrose. CTA intake suppression was characterized by reductions in burst size, average ingestion rate, and intraburst lick rate, and increases in brief pauses and burst counts. Compared with previous studies, LiCl licking shifted from a pattern initially matching that for normally accepted NaCl to one matching licking for normally avoided quinine hydrochloride by the end of the 1st acquisition trial. In Experiment 2, a novel paradigm was developed to show that rats expressed CTA generalization within 9 min of their first LiCl access. These results suggest that licking microstructure analysis can be used to assay changes in hedonic evaluation caused by treatments that produce aversive states.

Chorda Tympani Transection and Selective Desalivation Differentially Disrupt Two-Lever Salt Discrimination Performance in Rats

Water-restricted rats were trained to press 1 of 2 levers if a sampled stimulus was NaCl and the other lever if the stimulus was KCl (0.05, 0.1, or 0.2 M). Responses were reinforced with water. After training, the average rate of correct responses was 90%. Performance was unchanged following sham surgery. Chorda tympani (CT) transection reduced average discrimination performance to 67.7% correct, and extirpation of the sublingual and submaxillary salivary glands reduced average performance to 80% correct. Although selective desalivation moderately reduced discriminability, a disrupted salivary environment does not explain the effects of CT transection. More likely, the discrimination deficit in CT-transected rats reflects a loss of critical taste input conveyed by the CT about salts.

C57BL/6J and DBA/2J mice vary in lick rate and ingestive microstructure

Fluid licking in mice is an example of a rhythmic behavior thought to be under the control of a central pattern generator. Inbred strains of mice have been shown to differ in mean or modal interlick interval (ILI) duration, suggesting a genetic‐based variation. We investigated water licking in the commonly used inbred strains C57BL/6J (B6) and DBA/2J (D2), using a commercially available contact lickometer. Results from 20‐min test sessions indicated that D2 mice lick at a faster rate than B6 mice (10.6 licks/s vs. 8.5 licks/s), based on analysis of the distribution of short‐duration ILIs (50–160 ms). This strain difference was independent of sex, extent of water deprivation or total number of licks. D2 mice also displayed a faster lick rate when the strains were tested with a series of brief (5 s) trials. However, when ingestion over the entire 20‐min session was analyzed, it was evident that D2 mice had an overall slower rate of ingestion than B6 mice. This was because of the tendency for D2 mice to have more very long pauses (>30 s) between sequences of licking bursts. Overall, it appeared that D2 mice licked more efficiently, ingesting more rapidly during excursions to the spout that were fewer and farther between.

Combined Glossopharyngeal and Chorda Tympani Nerve Transection Elevates Quinine Detection Thresholds in Rats (Rattus norvegicus)

Using a conditioned shock avoidance procedure, behavioral quinine hydrochloride thresholds were measured before and after glossopharyngeal (GLX), chorda tympani (CTX), or combined glossopharyngeal and chorda tympani (GLX + CTX) transection, as well as after sham surgery. In Experiment 1, thresholds in the sham, CTX, and GLX rats (Rattus norvegicus) either improved (lowered) or remained the same after surgery. In Experiment 2, GLX + CTX caused a pronounced 1.5 log10 unit increase in presurgically measured thresholds. Neither the glossopharyngeal nor the chorda tympani nerve is necessary for normal sensitivity to low quinine concentrations provided the other is intact. When both of these nerve are transected, however, the remaining afferent input is not sufficient to maintain normal detection performance.

Glossopharyngeal Nerve Transection Reduces Quinine Avoidance in Rats Not Given Presurgical Stimulus Exposure

Behavioral studies on the effects of bilateral glossopharyngeal nerve (GL) transection on quinine responsiveness have yielded mixed results. These differences may be explained by the presence or absence of presurgical exposure with the tastant. In the present experiment we measured unconditioned licking to quinine in rats that had no exposure to quinine before surgery. Rats were water deprived and trained to lick water during 10 s trials in an automated gustometer. Next, they were divided into groups that received either GL transection or sham surgery (CON). Following recovery, the water-deprived rats were presented with seven concentrations of quinine hydrochloride (0.003-3 mM) and distilled water. The number of licks to each tastant was averaged over three days of testing. Rats with GL transection licked significantly more to the higher concentrations of quinine relative to CON rats, resulting in a 0.44 log10 unit shift in the quinine concentration-response curve. These results when considered with prior work suggest that experience before nerve transection may have a small protective effect on taste-guided behavioral responsiveness to quinine in rats.

Chorda Tympani Nerve Transection Disrupts Taste Aversion Learning to Potassium Chloride, but Not Sodium Chloride

In Experiment 1, rats with chorda tympani nerve transection (CTX) acquired a LiCl-conditioned taste aversion to 0.1 M NaCl at the same rate as controls. After 3 conditioning trials, the aversion generalized to 0.03 and 0.3 M NaCl, but did not generalize to KCI (0.03, 0.1, and 0.3 M), in either the sham or CTX group. In Experiment 2, the sham group, but not the CTX group, formed an aversion to 0.1 M KCI after 1 trial. The CTX rats did form a moderate aversion after 2 conditioning trials. Following the 3rd trial, the CTX group did not suppress licking to 0.03 or 0.3 M KCI or any concentration of NaCl in relation to controls. Although there is strong evidence that CTX affects NaCl taste perception, these findings indicate that, under certain conditions, rats can nonetheless distinguish NaCl from KCI after such neurotomy. Moreover, CTX appears to have a substantial effect on the perceived intensity of KCl.

Cellular expression of α-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, α‐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‐μm sections, which were incubated with antibodies against α‐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 α‐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 α‐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. J. Comp. Neurol. 409:118–130, 1999.