Taste responses of the gerbil IXth nerve

Summated taste responses to 12 taste solutions were recordedfrom the IXth (glossopharyngeal) nerve of 38 Mongolian gerbils(Meriones unguiculatus). 0.3 M NH₄Cl was the most effectivestimulant. The relative magnitude of the peak summated responsewas a positively accelerated function of log molar concentration.Absolute thresholds were determined for three chemicals: 0.002M NaCl, 0.0003 M HCl, and 0.002 M sucrose. The relative magnitudesof the responses to quinine, NH₄Cl, and KCl were greater forthe IXth nerve than for the chorda tympani nerve, whereas NaClwas more effective for the chorda tympani. A similar patternis seen in the rat. Acetic and citric acid may bind to commonreceptor sites. NH₄Cl, KCl, and HCl may also have receptor sitesin common.     

Greater superficial petrosal nerve transection in rats does not change unconditioned licking responses to putatively sweet taste stimuli

The greater superficial petrosal nerve (GSP), innervating taste buds in the palate, is known to be exceptionally responsive to sucrose, especially compared with the responsiveness of the chorda tympani nerve (CT). However, whereas transection of the CT (CTX) alone has little or no effect on unconditioned licking responses to many "sweet" stimuli, the impact of GSP transection (GSPX) alone is equivocal. To further examine the role of the GSP on licking responses to putatively sweet-tasting substances, brief-access taste tests were conducted in nondeprived rats before and after sham surgery (SHAM) or CTX or GSPX. A range of concentrations of sucrose, L-alanine, glycine, and L-serine, with and without 1.0 mM inosine monophosphate (IMP) added, were used. All groups showed significant concentration-dependent increases in licking to all stimuli presurgically and postsurgically. CTX decreased licking responses relative to SHAM rats in the first sucrose test. There was also a group x concentration interaction for L-alanine, but post hoc tests did not reveal its basis. Other than this, there were no significant differences among the surgical groups. Interestingly, rats with GSPX tended to initiate fewer trials than SHAM rats. Overall, after GSPX, the remaining gustatory nerves are apparently sufficient to maintain concentration-dependent licking responses to all stimuli tested here. The disparity between our results and others in the literature where GSPX reduced licking responses to sucrose is possibly related to differences in surgical technique or test trial duration.     

Effect of chorda tympani nerve transection on salt taste perception in mice

Effects of gustatory nerve transection on salt taste have been studied extensively in rats and hamsters but have not been well explored in the mouse. We examined the effects of chorda tympani (CT) nerve transection on NaCl taste preferences and thresholds in outbred CD-1 mice using a high-throughput phenotyping method developed in our laboratory. To measure taste thresholds, mice were conditioned by oral self-administration of LiCl or NaCl and then presented with NaCl concentration series in 2-bottle preference tests. LiCl-conditioned and control NaCl-exposed mice were given bilateral transections of the CT nerve (LiCl-CTX, NaCl-CTX) or were left intact as controls (LiCl-CNT, NaCl-CNT). After recovery from surgery, mice received a concentration series of NaCl (0-300 mM) in 48-h 2-bottle tests. CT transection increased NaCl taste thresholds in LiCl-conditioned mice and eliminated avoidance of concentrated NaCl in control NaCl-exposed mice. This demonstrates that in mice, the CT nerve is important for detection and recognition of NaCl taste and is necessary for the normal avoidance of high concentrations of NaCl. The results of this experiment also show that the method of high-throughput phenotyping of salt taste thresholds is suitable for detecting changes in the taste periphery in mouse genetic studies.     

Reinnervation of murine muscle following fetal sciatic nerve transection

A technique is reproted that permits transection of the sciatic nerve of mouse fetuses without interfering with fetal viability. Sciaticotomy was performed on Swiss Webster mice at day 17 of gestation; the contralateral side served as a control. Six weeks later the extensor digitorum longus (EDL) muscles on both sides were injected with horseradish peroxidase (HRP). Examination of the lumbar spinal cord revealed that while a substantial number of motor neurons in the region of the spinal cord giving rise to the sciatic nerve died, the EDL muscle did become reinnervated. The size of the EDL motor neuron pool on the denervated-reinnervated side was ～43% of that seen on the control side. While the control EDL motor neuron pool was located in lumbar segments L3–L5, the location of the pool to the denervated-reinnervated EDL was shifted cranially to L2–L4. Denervated-reinnervated EDL muscles were analyzed immunohistochemically to study the effect of fetal denervation on the neuronal cell adhesion molecule (N-CAM) expression. At 2 weeks postnatal, N-CAM immunoreactivity in control muscle was segregated to the motor end plate region, while fetally denervated muscle continued to express N-CAM along the length of the sarcolemma. Thus fetally denervated muscle does not develop the same pattern of N-CAM expression as normal, innervated muscle. By 6 weeks of age, the denervated-reinnervated muscle showed the same level and distribution of N-CAM immunoreactivity as did age-matched control muscle, indicating that most, if not all, of its myofibers had been reinnervated.     

Reinnervation of murine muscle following fetal sciatic nerve transection.

A technique is reported that permits transection of the sciatic nerve of mouse fetuses without interfering with fetal viability. Sciaticotomy was performed on Swiss Webster mice at day 17 of gestation; the contralateral side served as control. Six weeks later the extensor digitorum longus (EDL) muscles on both sides were injected with horseradish peroxidase (HRP). Examination of the lumbar spinal cord revealed that while a substantial number of motor neurons in the region of the spinal cord giving rise to the sciatic nerve died, the EDL muscle did become reinnervated. The size of the EDL motor neuron pool on the denervated-reinnervated side was approximately 43% of that seen on the control side. While the control EDL motor neuron pool was located in lumbar segments L3-L5, the location of the pool to the denervated-reinnervated EDL was shifted cranially to L2-L4. Denervated-reinnervated EDL muscles were analyzed immunohistochemically to study the effect of fetal denervation on the neuronal cell adhesion molecule (N-CAM) expression. At 2 weeks postnatal, N-CAM immunoreactivity in control muscle was segregated to the motor end-plate region, while fetally denervated muscle continued to express N-CAM along the length of the sarcolemma. Thus fetally denervated muscle does not develop the same pattern of N-CAM expression as normal, innervated muscle. By 6 weeks of age, the denervated-reinnervated muscle showed the same level and distribution of N-CAM immunoreactivity as did age-matched control muscle, indicating that most, if not all, of its myofibers had been reinnervated.     

Glossopharyngeal nerve transection does not alter taste reactivity to sucrose conditioned to be aversive

Glossopharyngeal nerve (GL) transection in rats is known to markedly reduce gaping, a stereotypical aversive oromotor behavior, in response to intraorally delivered quinine. In this experiment we tested whether GL transection would reduce gaping in response to an otherwise palatable stimulus (sucrose) but conditioned to be aversive. Sprague-Dawley rats were implanted with intraoral cannulae. Five received bilateral transection of the GL and five served as sham-operated controls. Water-deprived rats were presented with 0.3 M sucrose for 15 min immediately followed by an injection of 0.15 M LiCl on three occasions. Rats were then habituated to the taste reactivity chamber and intraoral fluid infusion for 3 days, and tested on day 4 with a 1 ml infusion (1 min) of 0.3 M sucrose. All rats drank negligible amounts of sucrose by the third conditioning session and there were no differences in sucrose intake between the groups. There were no significant differences in gapes, or any other measured oromotor response, to sucrose between GL-transected and sham-operated rats. These results show that the GL is not a necessary afferent limb for gaping in response to conditionally aversive taste compounds.     

Developmental changing of rat taste responses from chorda tympani nerve

In order to clarify developmental changing of gustatory system, histological and electrophysiological experiments were performed in the rat. Histological examination on the anterior tongue innervated by chorda tympani nerve showed that the ratio of matured taste buds which possess a taste pore were only 9% of all taste buds observed at 1 week of postnatal age, and 81.3% at 3 weeks of age. Recording integrated responses from the chorda tympani nerve reveals that taste buds with a pore at 1 week of age responded to NaCl, HCl, and quinine-HCl as well as in adult rats, which suggests that these relatively young taste buds are matured functionally for these three stimuli. However, the response magnitudes for various sugars at 1 week of age were smaller compared to those in the adult rat, reached to the maximum at 3 weeks of age, then decreased gradually with age. Also, results from the experiment of cross-adaptation among different sugars, effects of pronase-E treatment of the tongue, analysis of correlation between on- and off-responses to sugars, showed that qualitative changes for sugar responses continues after 3 weeks of age. These results suggest that functional changes occur in the gustatory processing of sugars during postnatal development in the rat chorda tympani nerve.     

Conduction velocity of the IXth nerve fibers innervating taste organs in the rostral and caudal tongue region in bullfrog

Sensory impulses elicited by electrical, mechanical and chemicalstimulation of the fungiform papillae in the bullfrog tonguewere recorded at two distant points on the glossopharyngealnerve, and the conduction velocities of different types of sensoryunits in the medial branch supplying the caudal two-thirds ofthe tongue were compared with those in the lateral branch innervatingthe rostral third. (1) Rapidly-and slowly-adapting mechanosensitiveunits in the medial branch had the mean conduction velocityof 23 ± 0.4 (S.E.) and 14.8 ± 0.3 m/sec, respectively,the former but not the latter being significantly (P <0.001)slower than the lateral branch units of the same type by 6m/sec.(2) The velocity of gustatory unit impulses was found to rangefrom 14 to 20 m/sec. Statistical examination revealed that themean conduction velocities of the medial branch units can bearranged in the order of water- > sucrose- > acetic acid-= NaCl- >quinine-sensitive units. Similar order had beenfound in the lateral branch units, which were significantly(P <0.001) faster in the mean conduction velocity than themedial branch units of the respective type by 2 m/sec.     

Changes in odor quality discrimination following recovery from olfactory nerve transection

Following recovery from olfactory nerve transection, animals regain their ability to discriminate between odors. Odor discrimination is restored after new neurons establish connections with the olfactory bulb. However, it is not known if the new connections alter odor quality perception. To address this question, 20 adult hamsters were first trained to discriminate between cinnamon and strawberry odors. After reaching criterion (> or = 90% correct response), half of the animals received a bilateral nerve transection (BTX) and half a surgical sham procedure. Animals were not tested again until day 40, a point in recovery when connections are re-established with the bulb. When BTX animals were tested without food reinforcement, they could not perform the odor discrimination task. Sham animals, however, could discriminate, demonstrating that the behavioral response had not been extinguished during the 40 day period. When reinforcement was resumed, BTX animals were able to discriminate between cinnamon and strawberry after four test sessions. In addition, their ability to discriminate between these two familiar odors was no different than that of BTX and sham animals tested with two novel odors, baby powder and coffee. These findings suggest that, after recovery from nerve transection, there are alterations in sensory perception and that restoration of odor quality discrimination requires that the animal must again learn to associate individual odor sensations with a behavioral response.      

The time course of taste bud regeneration after glossopharyngeal or greater superficial petrosal nerve transection in rats

We previously have published data detailing the time course of taste bud regeneration in the anterior tongue following transection of the chorda tympani (CT) nerve in the rat. This study extends the prior work by determining the time course of taste bud regeneration in the vallate papilla, soft palate and nasoincisor ducts (NID) following transection of either the glossopharyngeal (GL) or greater superficial petrosal (GSP) nerve. Following GL transection in rats (n = 6 per time point), taste buds reappeared in the vallate papilla between 15 and 28 days after surgery, and returned to 80.3% of control levels (n = 12) of taste buds by 70 days postsurgery. The first appearance and the final percentage of the normal complement of regenerated vallate taste buds after GL transection resembled that seen previously in the anterior tongue after CT transection. However, in the latter case, regenerated taste buds reached asymptotic levels by 42 days after surgery, whereas within the time frame of the present study, a clear asymptotic return of vallate taste buds was not observed. In contrast to the posterior (and anterior) tongue, only 25% of the normal complement of palatal taste buds regenerated by 112 days and 224 days after GSP transection (n = 9). The difference in regenerative capacity might relate to the surgical approach used to transect the GSP. These experiments provide useful parametric data for investigators studying the functional consequences of gustatory nerve transection and regeneration.     

Effects of chorda tympani nerve anesthesia on taste responses in the NST

Human clinical and psychophysical observations suggest that the taste system is able to compensate for losses in peripheral nerve input, since patients do not commonly report decrements in whole mouth taste following chorda tympani nerve damage or anesthesia. Indeed, neurophysiological data from the rat nucleus of the solitary tract (NST) suggests that a release of inhibition (disinhibition) may occur centrally following chorda tympani nerve anesthesia. Our purpose was to study this possibility further. We recorded from 59 multi- and single- unit taste-responsive sites in the rat NST before, during and after recovery from chorda tympani nerve anesthesia. During anesthesia, average anterior tongue responses were eliminated but no compensatory increases in palatal or posterior tongue responses were observed. However, six individual sites displayed increased taste responsiveness during anesthesia. The average increase was 32.9%. Therefore, disinhibition of taste responses was observed, but infrequently and to a small degree in the NST At a subset of sites, chorda tympani-mediated responses decreased while greater superficial petrosal-mediated responses remained the same during anesthesia. Since this effect was accompanied by a decrease in spontaneous activity, we propose that taste compensation may result in part by a change in signal-to-noise ratio at a subset of sites.      

Mechanism of desensitization in taste responses: Prolongation of frog taste nerve responses to bitter substances by adapting the tongue to CaCl2 solution

• 1.1. The taste nerve responses to bitter substances declined rapidly to the spontaneous level during stimulation, but were remarkably prolonged when the tongue was stimulated after adapting to CaCl₂ solution.
• 2.2. The prolongation of the responses (inhibition of desensitization) to bitter substances was not inhibited by the Ca-channel blockers, suggesting that the inhibition of the desensitization occurs not inside taste cells but at membrane level.

     

Ingestive responses to some heavy metal salts in mice and inhibition of taste nerve responses by metals

Preference-aversion for heavy metals in ddy mice was examinedusing the two-bottle preference technique. Mice preferred MnCl₂,CoCl₂ at certain concentration ranges, but avoided NiCl₂, CuCl₂,ZnCl₂ and CdCl₂. The avoidance threshold for the salts becamelower in the order of Mn > Co > Ni > Cd > Cu >Zn, though the degree of rejection for CuCl₂ was larger thanthat for ZnCl₂. Preferences for sucrose and Na saccharin werereduced by low concentrations of CuCl₂ or ZnCl₂. However, preferencefor glycine was scarcely affected by addition of either saltat 1 mM. Electrophysiological experiments in mice on the inhibitionby heavy metal salts of chords tympani nerve responses to thefour basic taste stimuli revealed that CuCl₂ and ZnCl₂ at 1mM inhibited the response to sucrose strongly and those to guinineHCl (QHCl) and NaCl moderately, but had no effect on the HClresponse. On the other hand, MnCl₂, CoCl₂ and NiCl₂ at 1 mMscarcely inhibited the response to sucrose but suppressed thoseto other stimuli. CoCl₂ moderately inhibited responses to allthe stimuli. In general, metal salts producing a larger inhibitionof neural responses to sucrose and QHCL are avoided by miceat a lower concentration.     

Time course of morphological alterations of fungiform papillae and taste buds following chorda tympani transection in neonatal rats

The time course of structural changes in fungiform papillae was analyzed in rats that received unilateral chorda tympani nerve transection at 10 days of age. Morphological differences between intact and denervated sides of the tongue were first observed at 8 days postsection, with an increase in the number of fungiform papillae that did not have a pore. In addition, the first papilla with a filiform-like appearance was noted on the denervated side at 8 days postsectioning. By 11 days after surgery, the total number of papillae and the number of papillae with a pore were significantly lower on the transected side of the tongue as compared to the intact side. At 50 days postsection, there was an average of 70.5 fungiform papillae on the intact side and a mean of only 20.8 fungiform papillae the denervated side. Of those few remaining papillae on the cut side, an average of 13.5 papillae were categorized as filiform-like, while no filiform-like papillae occurred on the intact side. Significant reduction in taste bud volume was noted at 4 days posttransection and further decrements in taste bud volume were noted at 8 and 30 days postsection. Electron microscopy of the lingual branch of the trigeminal nerve from adult rats that received neonatal chorda tympani transection showed normal numbers of both myelinated and unmyelinated fibers. Thus, in addition to the well-characterized dependence of taste bud maintenance on the chorda tympani nerve, the present study shows an additional role of the chorda tympani nerve in papilla maintenance during early postnatal development.     

Anion modulation of taste responses in sodium-sensitive neurons of the hamster chorda tympani nerve

Beidler's work in the 1950s showed that anions can strongly influence gustatory responses to sodium salts. We have demonstrated "anion inhibition" in the hamster by showing that the chorda tympani nerve responds more strongly to NaCl than to Na acetate over a wide range of concentrations. Iontophoretic presentation of Cl- and acetate to the anterior tongue elicited no response in the chorda tympani, suggesting that these anions are not directly stimulatory. Drugs (0.01, 1.0, and 100 microM anthracene-9-carboxylate, diphenylamine-2-carboxylate, 4- acetamido-4'-isothiocyanatostilbene-2,2'-disulfonate, and furosemide) that interfere with movements of Cl- across epithelial cells were ineffective in altering chorda tympani responses to 0.03 M of either NaCl or Na acetate. Anion inhibition related to movements of anions across epithelial membranes therefore seems unlikely. The chorda tympani contains a population of nerve fibers highly selective for Na+ (N fibers) and another population sensitive to Na+ as well as other salts and acids (H fibers). We found that N fibers respond similarly to NaCl and Na acetate, with spiking activity increasing with increasing stimulus concentration (0.01-1.0 M). H fibers, however, respond more strongly to NaCl than to Na acetate. Furthermore, H fibers increase spiking with increases in NaCl concentration, but generally decrease their responses to increasing concentrations of Na acetate. It appears that anion inhibition applies to taste cells innervated by H fibers but not by N fibers. Taste cells innervated by N fibers use an apical Na+ channel, whereas those innervated by H fibers may use a paracellularly mediated, basolateral site of excitation.     

Rat taste nerve responses to salts carrying cations of large molecular size; are the taste responses to the salts induced by cation transport across apical membranes of taste cells?

1. The responses of rat chorda tympani nerve to various salts carrying cations of large molecular size which have small permeability were measured. 2. Salts carrying polyvalent cations such as Fe3+ or La3+ elicited much larger responses than NaCl or KCl. 3. Ammonium chloride derivatives having methyl or ethyl groups and salts carrying other organic cations of large molecular size elicited the responses comparable to that induced by NH4Cl or NaCl. 4. It was suggested that the taste responses to the salts carrying the cations of large molecular size are induced not by the cation transport but by adsorption of the cations on the membranes.