Multiple sensitivity of single taste cells of the frog tongue to four basic taste stimuli

The electrical response of the taste cells of the frog fungiform papillae to four fundamental taste solutions (NaCl, acetic acid, quinine-HCl and sucrose) was studied by using the intracellular recording technique. The average value of resting membrane potential was 22.5 mV, inside negative. Each of the four taste solutions applied to the tongue produced a slow depolarizing potential, the receptor potential, on which no spike potential was superimposed. The amplitude of the receptor potentials increased linearly as a function of the logarithm of the concentration of the stimulus. Amplitudes of depolarizations to a given taste stimulation varied from one cell to another even within a single taste bud. Most of the cells responded to more than two of the four basic taste solutions. Sensitivity patterns in terms of the number of effective solutions and the relative effectiveness of different kinds of solutions were variable among cells. Statistical analysis suggests that at the receptor membranes of the taste cells, the sensitivities for the four basic stimuli are independent and random.     

Centrifugal decrement in diameter of myelinated afferent fibres innervating frog taste organ

1. Regional changes in the diameter of single myelinated afferent nerve fibres innervating the taste disc of the fungiform papillae on the bullfrog tongue were investigated morphologically and functionally. 2. The diameter of myelinated afferents in the medial lingual branch of the glossopharyngeal nerve averaged 8.4 microns at the proximal end of the tongue and gradually decreased at the rate of 0.8 micron/cm length of the fibres as they ran in the apical direction of the tongue. 3. The conduction velocity of single myelinated afferent fibres within the tongue decreased gradually as they ran peripherally. 4. Electrophysiological inspection of neural connections between the fungiform papillae suggests that a gradual centrifugal decrease in the diameter of a single myelinated afferent fibre is not due to multiple bifurcations of the fibre at various sites within the tongue, but due to a natural gradual decrease in the thickness of the myelin sheath and the diameter of axon.     

Off-response in frog taste nerve and cell after stimulation of the tongue with bitter solutions

• 1.1. After a Ringer-adapted frog tongue was stimulated with quinine-HCl (Q-HCl), rinsing the tongue with the Ringer produced a large off-response in the glossopharyngeal nerve.
• 2.2. The off-response was caused by the enhancing action of Q-HCl stimulation upon the stimulating effectiveness of an NaCl component of Ringer solution.
• 3.3. Analysis of gustatory neural unit responses showed that following Q-HCl stimulation the enhancement of responses to Ringer of those units which responded to both Q-HCl and Ringer or Ringer alone is related to the generation of the off-response.
• 4.4. A phasic off-depolarization of taste cells elicited by a Ringer rinse following Q-HCl stimulation is thought to be associated with the off-response in the gustatory nerve.

     

Arterial perfusion of frog tongue for intracellular recording of taste cell receptor potential

The frog tongue was perfused through its artery with a Ringer solution using a peristaltic pump, and a method was developed to record stable intracellular receptor potentials of taste cells. Perfusing at 0.05 ml/min with a Ringer solution containing 5% dextran did not cause tongue edema, but perfusing at the same rate with Ringer without dextran caused edema. After perfusion at 0.05 ml/min with 100 mM K Ringer, the membrane potential of taste cells gradually decreased and reached a constant level in about 30 min, indicating that the intercellular fluid of the tongue could be replaced within this time period. While the artery of the frog tongue was perfused at 0.05 ml/min with Ringer containing 5% dextran, intracellular receptor potentials of taste cells elicited by four basic taste stimuli (1 M NaCl, 10 mM quinine-HCl (Q-HCl), 1 mM acetic acid and 1 M galactose) were similar to those obtained from the control taste cells under normal blood flow.     

Electrical responses of supporting cells in the frog taste organ to chemical stimuli

1. The mean resting potential of supporting cells in the frog taste organ was -19.1 mV. The supporting cells responded to the four basic taste stimuli with a depolarization but responded to water with a depolarization or a hyperpolarization. 2. The membrane resistances of supporting cells decreased during stimulation with sucrose, NaCl and acetic acid, but increased during stimulation with Q-HCl and water. 3. Reversal potential of the depolarizing response for 0.5 M NaCl in supporting cells was +7.6 mV. The depolarizing responses for Q-HCl and acetic acid were independent of the membrane potential level. 4. These results suggest that the characteristics of taste responses in supporting cells are similar to those in taste cells.     

A fluorescence histochemical study of the monoamine-containing cell in the developing frog taste organ

Summary Sequential changes in the monoamine-contianing cell (MC cell) of the developing frog tongue has been studied by fluorescence histochemistry using uptake of 5,6-dihydroxytryptamine. At st. 16, a few yellow fluorescent cells, here called MC cells, appeared in random order at the uppermost layer of the dorsal epithelium. They were round or elliptical in shape. At st. 18 the MC cells, greatly transformed, were found at the periphery of the sensory disc primordium which first appears during this stage. The MC cell was made up of three parts: perikaryon, process and terminal portion. The perikaryon was located at the upper half of the epithelium and from it a single process stretched vertically toward the basal lamina, above which the dilated terminal portion was found. Thereafter the perikaryon gradually moved toward the basal layer while remaining at the periphery of the disc primordium. Meanwhile the terminal portion moved over the basal lamina toward the center of the disc primordium. At st. 22, the whole of the MC cell lay flat above the basal lamina. The perikaryon was localized at the periphery of the sensory disc and from there the process stretched toward the center. Thus, the morphology of MC cells resembled the adult state, except for smaller size. MC cells were never observed in the subepithelial connective tissue in the present study. This seems to suggest that the MC cell of the frog fungiform papilla is of epithelial origin.     

Effects of antidromic activity in gustatory nerve fibers on taste disc cells of the frog tongue

Summary Antidromic electrical stimulation of the lingual branch of the glossopharyngeal (IX) nerve of the frog was carried out while recording intracellular potentials of taste disc cells.Antidromic activation of sensory fibers resulted in depolarization of cells of the upper layer of the disc and most commonly in hyperpolarization of the cells in the lower layer. These changes in potential exhibited latencies greater than 1 s (Fig. 3), and thus cannot be due to electrotonic effects of action potentials in terminals of IX nerve fibers, which have much shorter conduction times. These cell potentials also showed summation, adaptation and post-stimulus rebound (Figs. 3, 4).Depending upon the chemical stimulus used, antidromic activity produced either depression or enhancement of gustatory fiber discharge in response to taste stimuli (Fig. 5).Alteration of the resting membrane potential by current injection did not significantly modify the antidromically evoked potentials (Fig. 8), whereas chemical stimulation of the tongue did (Fig. 7), indicating that these potential changes are not the result of passive electrical processes.These experimental results indicate that the membrane potential of taste disc cells can be modified by antidromic activity in their afferent nerves. This mechanism may be responsible for peripheral interactions among gustatory units of the frog tongue.The research was supported in part by NIH grant NS-09168.     

Immunohistochemical investigation of the nitrergic system in the taste organ of the frog,Rana esculenta

We have studied by immunocytochemistry, the taste discs of the frog, Rana esculenta, with the aim of providing morphological and neurochemical data on the nitrergic system and of assessing the eventual presence of intrinsic neurons associated with the gustatory organs. In taste discs, antibodies against neuronal nitric oxide synthase (nNOS) revealed a positive immunoreaction in the taste receptor cell bodies and processes. The basal cells were also stained. All the fungiform papillae contained intragemmal nerve fibers showing nNOS immunoreactivity; these fiber were mainly located in the basal plexus. Immunoreactive nerve fibers were also visible at the periphery of the papilla-contacting ciliate cells, which form a ring around the taste disc. In conclusion, the findings obtained in this study suggest that the occurrence of nNOS-immunoreactivity in basal cells, taste cells and nerves might reflect a role for nitric oxide in taste mechanisms of Amphibia. The results may also sustain the physiological implication of NO as a molecule involved in the local target function of maintaining the taste bud mucosal integrity and in regulating the blood flow to the epithelium.     

Phenylthiocarbamide taste sensitivity and its relationship to growth variation

The hypothesis that there is a relationship between the tasting polymorphism and growth variation was tested on a sample of 425 Negro elementary school children. Twenty-seven non-tasters were found by testing with impregnated papers; 13 were male, 14 female, indicating no sex differences in this group. Matchedpair comparisons indicated no differences in weight, a tendency for tasters to be taller, and a stronger tendency for tasters of both sexes to be skeletally more mature. It was felt that the tendency for tasters to be taller might reflect their more advanced maturation status. The relationship to skeletal maturation might be indicative of the decreased thryoid activity found in other studies of phenylthiocarbamide tasting.     

Taste performance on the anterior human tongue varles with fungiform taste bud density

This study demonstrated that individual diversities of tastesensitivity on the anterior tongue are due, in part, to variationsin fungiform taste bud density. Citric acid solutions were deliveredto a closed, spatially-matched (each test site was 43 mm²) flowchamber attached to the surface of the anterior tongue in 84subjects. A two-alternative forced choice, modified staircaseprocedure was used to derive a detection threshold value forcitric acid. The same session also included a visual analograting procedure to scale the taste intensity judgement of fiveconcentrations of citric acid. The taste buds within the chamberwere distinguished by methylene blue stain and recorded by videomicroscopy.The sip-and-spit method was used to contrast the spatially-matchedcondition with whole mouth stimulation. We found that detectionthreshold values were inversely related to the number of fungiformtaste buds, independent of gender or age. Whole-mouth thresholdvalues determined for each subject were always well below thespatially-matched threshold values. In addition, the data demonstratedthat subjects were able to scale the dynamic range of citricacid solutions. However, y-intercept approached zero intensityas the number of fungiform papillae decreased. The observeddifferences in citric acid sensitivity and fungiform taste buddensity indicated that taste performance on the anterior tonguein humans varies, in part, with the number of taste buds.     

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.

     

Extraversion and taste sensitivity

The rationale for investigating the gustatory reactivity as influenced by personality dimensions was suggested by some prior findings of an association between extraversion and acuity in other sensory systems. Detection thresholds for sweet, salty, and bitter qualities of taste were measured in 60 young healthy male and female volunteers using a two-alternative forced-choice technique. Personality of the responders was assessed using the Eysenck Personality Inventory. Multivariate analysis of variance failed to demonstrate a statistically significant interaction between an extraversion-introversion score, neuroticism score, smoking, gender and age. The only reliable negative association was found between the body mass index (BMI) and taste sensitivity (Roy's largest root = 0.05, F(7436.5) = 8.34, P = 0.003). Possible reasons for lack of differences between introverts and extraverts in the values of taste detection thresholds were discussed.     

Structure and physiological properties of the taste organs on the ventral side of frog tongue (Rana catesbeiana)

The structure and physiological properties of the non-papillarytaste organs on the ventral side of frog tongue were examined.The taste organs were distributed around the hyoglossal muscleof the tongue. The total number of organs was estimated as about100. The fine structure of the taste organs was similar to thatof the fungiform papilla. Neural responses recorded from theglossopharyngeal nerve by applying chemical stimuli to the ventraltide of the tongue were noticeably smaller than those to thedorsal side of the tongue. The taste cells on the ventral sideof the tongue generated receptor potentials to various chemicalstimuli like those on the dorsal side of the tongue.     

Effect of sensory and sympathetic denervation of the frog tongue on the catecholamine containing cells of the taste buds]

The structure of catecholamine-containing dumb-bell shaped cells of the taste buds was studied by luminescent microscopy in the epithelial layer of the frog's tongue (Rana temporaria). On the unilateral section of the lingual nerve, a maintained adrenergic innervation of vessels and of the epithelium was observed, a decreased number of dumb-bell shaped cells in the taste bud, and their significant enlargement, and increased cathecholamine luminescence. With desympathization, no adrenergic nerves were observed on the vessels and the epithelium of the tongue. The size of the taste buds in desympathized cells of the tongue is sharply decreased and their number is increased. There is a tendency to grouping of the dumbbell shaped cells into 3--4 taste buds in one fungiform papillina. The experiments with sensory and sympathetic denervation of the frog tongue distinctly showed the trophic action of sensory and sympathetic nerves on the taste organ of the frog.     

Development of the tongue and taste disks in Pelobates fuscus

In the tadpole of Pelobates fuscus the process of tongue formation starts at the 32nd developmental stage. In more advanced stages (older than 38th) fast anterior and faucial growth of the tongue fold has been observed. This process is accompanied by the development of the gustatory organs. The dorsal surface of the tongue fold, smooth at the beginning, in older tadpoles (developmental stages 36-39th) forms protrusions in which gustatory organs of the taste disk type (TDs) develop. In the 41 st tadpole developmental stage anlages of TDs are formed by elongated cells, located more or less perpendicularly to the surface of the tongue. The diameter of the sensory area of a TD at the 45th developmental stage amounts to 94 microm, while in metamorphosed individuals it reaches 130-140 microm. At the base of a TD the presence of basal cell morphologically similar to that of Merkel cell was observed at the 42nd developmental stage of a tadpole. Fully developed afferent synaptic connections in the sensory epithelium of a TD were found starting from the 44th developmental stage. Single synaptic vesicles with an electron-dense core were observed in gustatory cells as early as at the 41 st developmental stage of the tadpole. From the observations reported here it can be inferred that in Pelobates fuscus development of both the tongue and TDs is similar to that already described in the representatives of the Rana genus.     

The ultrastructure of taste and touch receptors of the Frog's taste organ

Summary The taste buds from fungiform papillae and the hard palate of frogs were investigated with the scanning and transmission electron microscopes. An immature pre-taste cell and a mature taste cell can be differentiated. Only the mature taste cell exhibits synaptic contact with the afferent taste fibre. Glandular and satellite supporting cells envelop the thin apical processes of the sensory cells. At the base of the taste disc up to 10 Merkel cells form a complex with nerve endings. There are two types of myelinated fibres, large and small. The small fibre innervates the taste cells, the thicker nerve fibre the Merkel cells. The occurrence of two types of receptors explains physiological results.Supported by the Deutsche Forschungsgemeinschaft Rezeptorphysiologie.