Depression of gustatory receptor potential in frog taste cell by parasympathetic nerve-induced slow hyperpolarizing potential

Parasympathetic nerve (PSN) innervates taste cells of the frog taste disk, and electrical stimulation of PSN elicited a slow hyperpolarizing potential (HP) in taste cells. Here we report that gustatory receptor potentials in frog taste cells are depressed by PSN-induced slow HPs. When PSN was stimulated at 30 Hz during generation of taste cell responses, the large amplitude of depolarizing receptor potential for 1 M NaCl and 1 mM acetic acid was depressed by approximately 40% by slow HPs, but the small amplitude of the depolarizing receptor potential for 10 mM quinine-HCl (Q-HCl) and 1 M sucrose was completely depressed by slow HPs and furthermore changed to the hyperpolarizing direction. The duration of the depolarizing receptor potentials depressed by slow HPs prolonged with increasing period of PSN stimulation. As tastant-induced depolarizing receptor potentials were increased, the amplitude of PSN-induced slow HPs inhibiting the receptor potentials gradually decreased. The mean reversal potentials of the slow HPs were approximately -1 mV under NaCl and acetic acid stimulations, but approximately -14 mV under Q-HCl and sucrose stimulations. This implies that when a slow HP was evoked on the same amplitude of depolarizing receptor potentials, the depression of the NaCl and acetic acid responses in taste cells was larger than that of Q-HCl and sucrose responses. It is concluded that slow HP-induced depression of gustatory depolarizing receptor potentials derives from the interaction between gustatory receptor current and slow hyperpolarizing current in frog taste cells and that the interaction is stronger for NaCl and acetic acid stimulations than for Q-HCl and sucrose stimulations.     

Electrical properties and gustatory responses of various taste disk cells of frog fungiform papillae

We compared the electrical properties and gustatory response profiles of types Ia cell (mucus cell), Ib cell (wing cell), and II/III cell (receptor cell) in the taste disks of the frog fungiform papillae. The large depolarizing responses of all types of cell induced by 1 M NaCl were accompanied by a large decrease in the membrane resistance and had the same reversal potential of approximately +5 mV. The large depolarizing responses of all cell types for 1 mM acetic acid were accompanied by a small decrease in the membrane resistance. The small depolarizing responses of all cell types for 10 mM quinine-HCl (Q-HCl) were accompanied by an increase in the membrane resistance, but those for 1 M sucrose were accompanied by a decrease in the membrane resistance. The reversal potential of sucrose responses in all cell types were approximately +12 mV. Taken together, depolarizing responses of Ia, Ib, and II/III cells for each taste stimulus are likely to be generated by the same mechanisms. Gustatory depolarizing response profiles indicated that 1) each of Ia, Ib, and II/III cells responded 100% to 1 M NaCl and 1 mM acetic acid with depolarizing responses, 2) approximately 50% of each cell type responded to 10 mM Q-HCl with depolarizations, and 3) each approximately 40% of Ia and Ib cells and approximately 90% of II/III cells responded to 1 M sucrose with depolarizations. These results suggest that the receptor molecules for NaCl, acid, and Q-HCl stimuli are equivalently distributed on all cell types, but the receptor molecules for sugar stimuli are richer on II/III cells than on Ia and Ib cells. Type III cells having afferent synapses may play a main role in gustatory transduction and transmission.     

Interaction Between Gustatory Depolarizing Receptor Potential and Efferent-Induced Slow Depolarizing Synaptic Potential in Frog Taste Cell

Electrical stimulation of parasympathetic nerve (PSN) efferent fibers in the glossopharyngeal nerve induced a slow depolarizing synaptic potential (DSP) in frog taste cells under hypoxia. The objective of this study is to examine the interaction between a gustatory depolarizing receptor potential (GDRP) and a slow DSP. The amplitude of slow DSP added to a tastant-induced GDRP of 10 mV was suppressed to 60% of control slow DSPs for NaCl and acetic acid stimulations, but to 20–30% for quinine–HCl (Q-HCl) and sucrose stimulations. On the other hand, when a GDRP was induced during a prolonged slow DSP, the amplitude of GDRPs induced by 1 M NaCl and 1 M sucrose was suppressed to 50% of controls, but that by 1 mM acetic acid and 10 mM Q-HCl unchanged. It is concluded that the interaction between GDRPs and efferent-induced slow DSPs in frog taste cells under hypoxia derives from the crosstalk between a gustatory receptor current across the receptive membrane and a slow depolarizing synaptic current across the proximal subsynaptic membrane of taste cells.     

Membrane resistance change of the frog taste cells in response to water and Nacl

The electrical properties of the frog taste cells during gustatory stimulations with distilled water and varying concentrations of NaCl were studied with intracellular microelectrodes. Under the Ringer adaptation of the tongue, two types of taste cells were distinguished by the gustatory stimuli. One type, termed NaCl-sensitive (NS) cells, responded to water with hyperpolarizations and responded to concentrated NaCl with depolarizations. In contrast, the other type of cells, termed water-sensitive (WS) cells, responded to water depolarizations and responded to concentrated NaCl with hyperpolarizations. The membrane resistance of both taste cell types increased during the hyperpolarizing receptor potentials and decreased during the depolarizing receptor potentials, Reversal potentials for the depolarizing and hyperpolarizing responses in each cell type were a few millivolts positive above the zero membrane potential. When the tongue was adapted with Na-free Ringer solution for 30 min, the amplitude of the depolarizing responses in the NS cells reduced to 50% of the control value under normal Ringer adaptation. On the basis of the present results, it is concluded (a) that the depolarizing responses of the NS and WS cells under the Ringer adaptation are produced by the permeability increase in some ions, mainly Na+ ions across the taste cell membranes, and (b) that the hyperpolarizing responses of both types of taste cells are produced by a decrease in the cell membrane permeability to some ions, probably Na+ ions, which is slightly enhanced during the Ringer adaptation.     

Topographical difference in taste organ density and its sensitivity of frog tongue

Distribution density of the taste disks of the fungiform papillae in the frog tongue was larger at the proximal portion than at the apical and middle portions. The number of myelinated afferent nerve fibres and taste cells per cm2 area of the tongue increased in the order of proximal greater than middle greater than apical portion. The amplitudes of gustatory neural responses for 0.5 M NaCl, 0.5 M KCl, 0.5 M NH4Cl, 0.05 M CaCl2, 1 mM acetic acid and 1 mM quinine-HCl (Q-HCl) were significantly larger with lingual stimulation of the proximal region than with the stimulation of the apical region. With these stimuli the mean ratio of the apical response to the proximal response was 1.00:1.54. On the other hand, this ration with deionized water was 1.00:5.00. The mean magnitudes of receptor potentials in taste cells for 1 mM acetic acid and 10 mM Q-HCl were the same among the apical, middle and proximal portions of the tongue. The mean magnitudes of receptor potentials for 0.5 M NaCl were significantly larger at the apical portion than at the other portions, whereas those for deionized water tended to be the largest at the proximal portion. It is concluded that the larger magnitude of the gustatory neural responses at the proximal portion of the tongue is due to morphological and physiological properties of the taste organ.     

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.     

Non-synaptic transformation of gustatory receptor potential by stimulation of the parasympathetic fiber of the frog glossopharyngeal nerve

When the glossopharyngeal nerve (GP) in the frog was strongly stimulated electrically, slow potentials were elicited from the tongue surface and taste cells in the fungiform papillae. Injection of atropine completely blocked these slow potentials. The present and previous data indicate that the slow potentials induced in the tongue surface and taste cells are due to a liquid junction potential between saliva secreted from the lingual glands due to parasympathetic fiber activity and an adapting solution on the tongue surface. Intracellularly recorded depolarizing receptor potentials in taste cells induced by 0.5 M NaCl and 3 mM acetic acid were enhanced by depolarizing slow potentials induced by GP nerve stimulation, but were depressed by the hyperpolarizing slow potentials. On average, the receptor potential of taste cells for 0.5 M NaCl was increased by 25% by the GP nerve-induced slow potential, but the receptor potential of taste cells for 3 mM acetic acid was decreased by 1% by the slow potential. These transformations of receptor potentials in frog taste cells were not due to a synaptic event initiated between taste cells and the efferent nerve fiber, but due to a non-synaptic event, a lingual junction potential generated in the dorsal lingual epithelium by GP nerve stimulation.     

Dye-coupling among frog (Rana catesbeiana) taste disk cells

• 1.1. Dye-coupling among taste disk cells in the bullfrog fungiform papillae was examined histologically by injecting a fluorescent dye (Lucifer yellow) into the cell, and the effects of the dye-coupling on depolarizing responses induced by taste stimuli were studied electrophysiologically.
• 2.2. With dye injection into a taste cell, dye-coupling was found between taste cells (23%) or between taste cell and supporting cell (28%). With dye injection into a supporting cell, dye-coupling was found between supporting cells (34%) or between supporting cell and taste cell (27%).
• 3.3. Depolarizing responses recorded from either a taste cell or a supporting cell to stimulation with 0.5 M NaCl or 10 mM quinine-HCl were the same in amplitude whether the dye-coupling to another cell was present or not. On the other hand, depolarizing responses recorded from a taste cell for 0.5 mM acetic acid became significantly larger when dye-coupled to a supporting cell.
• 4.4. It is concluded that gustatory transduction for acid stimuli is influenced by supporting cells coupled to taste cells.

     

OFF-RESPONSE OF GUSTATORY NERVE FOLLOWING TERMINATION OF QUININE STIMULI APPLIED TO THE FROG TONGUE

After a Ringer-adapted frog tongue was stimulated with 0.001M quinine-HCl(Q-HCl) in deionized water, the Ringer rinse ofthe tongue elicited a large gustatory nerve response. Sincethe Ringer-adapted tongue did not respond to Ringer solution,this nerve response after Q-HC1 is termed an off-response. Itwas revealed that the off-response of gustatory nerve to theRinger rinse resulted from the enhancing effect of Q-HCl adaptationupon the response to 111.2 mM NaCl component of Ringer whichwas ineffective for Ringer-adapted tongue. Weaker enhancementof the response to NaCl of Ringer was also produced by adaptingthe tongue to water. Therefore, the enhancing effect of Q-HClin deionized water is the summed effect of Q-HCl solute andwater solvent. Microelectrode study revealed that during theadaptation to Q-HCl the membrance potential of some NaCl-sensitivetaste cells was more hyperpolarized than that of Ringer-adaptedstate. The larger membrane potential maintained under Q-HCladaptation was markedly depolarized by the NaCl component ofthe Ringer. This sufficient depolarization response of the tastecells might be associated with generation of the off-responsein the gustatory nerve.     

Membrane potential responses of paramecium caudatum to external Na+

The membrane potential responses of Paramecium caudatum to Na+ ions were examined to understand the mechanisms underlying the sensation of external inorganic ions in the ciliate by comparing the responses of the wild type and the behavioral mutant. Wild-type cells exhibited initial continuous backward swimming followed by repeated transient backward swimming in the Na+-containing test solution. A wild-type cell impaled by a microelectrode produced initial action potentials and a sustained depolarization to an application of the test solution. The prolonged depolarization, the depolarizing afterpotential, took place subsequently after stimulation. The ciliary reversal of the cell was closely associated with the depolarizing responses. When the application of the test solution was prolonged, the wild-type cell produced sustained depolarization overlapped by repeated transient depolarization. A behavioral mutant defective in the Ca2+ channel, CNR (caudatum non reversal), produced a sustained depolarization but no action potential or depolarizing afterpotential. The mutant cell responded to prolonged stimulation with sustained depolarization overlapped by transient depolarization, although it did not show backward swimming. The results suggest that Paramecium shows at least two kinds of membrane potential responses to Na+ ions: a depolarizing afterpotential mediating initial backward swimming and repeated transient depolarization responsible for the repeated transient backward swimming.     

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.

     

The adaptation of the frog tongue to bitter solutions: Enhancing effect on gustatory neural response to salt stimuli

• 1.1. The initial phasic component of frog gustatory neural responses to various 0.1 M salt solutions was greatly augmented in amplitude after the tongue was adapted for 10 sec to 1 mM quinine-HC1 (Q-HCl), quinine-H₂SO₄ (Q-H₂SO₄) and picric acid.
• 2.2. Out of 103 examined gustatory units responding to both 0.5 M NaCl and 1 mM Q-HCl or to 0.5 M NaCl alone, 67% exhibited an enhancement of response to the NaCl after the Q-HCl adaptation but the remaining 33% showed a suppression or no alteration of NaCl response after the Q-HCl.
• 3.3. Intracellular taste cell responses to salt stimuli after 1 mM Q-HCl adaptation showed an initial phasic depolarization which was not observed under control Ringer adaptation. This de polarization might be concerned with the enhancement of initial phasic neural responses to salts following the Q-HCl adaptation.

     

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.     

Vasopressin increases frog gustatory neural responses elicited by NaCl and HCl.

1. The effect of arginine vasopressin (AVP) on frog gustatory responses was investigated by recording integrated responses of the whole glossopharyngeal nerve by stimulation of the tongue with tastants. 2. After AVP (100 mUnits/ml) was perfused to the basolateral side of taste cells through the lingual artery, gustatory neural responses for NaCl and hydrochloric acid (HCl) stimuli were greatly enhanced, but the responses for CaCl2, quinine hydrochloride (Q-HCl) and galactose were not affected. 3. Three hours after the onset of AVP perfusion, the responses for NaCl and HCl increased to 260% and 270% of the respective controls. 4. The NaCl response which was insensitive to amiloride during normal saline perfusion became sensitive to amiloride during AVP perfusion. 5. When membrane-permeable 8-bromo-cyclic AMP (8-Br-cAMP, 0.1 mM) was perfused to the basolateral side of taste cells, the responses for NaCl and HCl decreased to 41 and 63% of the respective controls. 6. These results suggest that AVP may regulate the gustatory responses for monovalent salts and acids by a mechanism which is not necessary to activate adenylate cyclase.     

Response characteristics of rat taste cells to potassium benzoate

The rat taste cells responded to K-benzoate solutions higher than the threshold concentrations (0.03-0.3 M) with a depolarizing receptor potential, but they responded to K-benzoate lower than the thresholds with a hyperpolarizing receptor potential. In either depolarizing or hyperpolarizing receptor potentials the rise time decreased with increasing amplitude, but the fall time increased with increasing amplitude. During generation of either depolarizing or hyperpolarizing receptor potentials the input resistance of taste cells decreased with increasing amplitude. Application of the mixtures of various concentrations of NaCl and 0.05 M K-benzoate resulted in a reduction of receptor potential amplitude, as compared with that evoked by application of NaCl alone. It is concluded that a depression of gustatory neural impulse frequency by low concentrations of K-benzoate is mainly due to the hyperpolarizing receptor potential of taste cells elicited by the K-benzoate solutions.     

Membrane properties of isolated mudpuppy taste cells

The voltage-dependent currents of isolated Necturus lingual cells were studied using the whole-cell configuration of the patch-clamp technique. Nongustatory surface epithelial cells had only passive membrane properties. Small, spherical cells resembling basal cells responded to depolarizing voltage steps with predominantly outward K+ currents. Taste receptor cells generated both outward and inward currents in response to depolarizing voltage steps. Outward K+ currents activated at approximately 0 mV and increased almost linearly with increasing depolarization. The K+ current did not inactivate and was partially Ca++ dependent. One inward current activated at -40 mV, reached a peak at -20 mV, and rapidly inactivated. This transient inward current was blocked by tetrodotoxin (TTX), which indicates that it is an Na+ current. The other inward current activated at 0 mV, peaked at 30 mV, and slowly inactivated. This more sustained inward current had the kinetic and pharmacological properties of a slow Ca++ current. In addition, most taste cells had inwardly rectifying K+ currents. Sour taste stimuli (weak acids) decreased outward K+ currents and slightly reduced inward currents; bitter taste stimuli (quinine) reduced inward currents to a greater extent than outward currents. It is concluded that sour and bitter taste stimuli produce depolarizing receptor potentials, at least in part, by reducing the voltage-dependent K+ conductance.     

Characteristics of biphasic slow depolarizing and slow hyperpolarizing potential in frog taste cell induced by parasympathetic efferent stimulation

When the velocity of capillary blood flow in the frog tongue declined to an intermediate range of 0.2-0.7 mm/s, the glossopharyngeal nerve stimulation induced a biphasic slow depolarizing and slow hyperpolarizing potential (HP) in taste cells. The objective of this work was to examine the generative mechanisms of the biphasic slow potentials. The biphasic slow response was always preceded by a slow depolarizing potential (DP) component and followed by a slow HP component. Intravenous injection of tubocurarine completely blocked the biphasic slow responses, suggesting that both components of the biphasic slow potentials are evoked by the parasympathetic nerve (PSN) fibers. Membrane conductance of taste cells increased during slow DPs and decreased during slow HPs. The reversal potential of either component of a biphasic slow response was the almost same value of -12 mV. An antagonist, L-703,606, for neurotransmitter substance P neurokinin(1) receptor completely blocked both components of the biphasic slow responses. An antagonist, flufenamic acid, for nonselective cation channels on the taste cell membrane completely blocked the biphasic slow responses. These results suggest that PSN-induced biphasic slow responses are postsynaptically elicited in taste cells by releasing substance P at the PSN axon terminals. It is concluded that the slow DP component may be generated by opening one type of nonselective cation channel on taste cells and that the slow HP component may be generated by closing the other type of nonselective cation channel. We discussed that a second messenger inositol 1,4,5-trisphosphate might be related to a slow DP component and another second messenger diacylglycerol might be related to a slow HP component.     

Enhancement of Gustatory Neural Responses by Parasympathetic Nerve in the Frog

The autonomic nervous system affects the gustatory responses in animals. Frog glossopharyngeal nerve (GPN) contains the parasympathetic nerve. We checked the effects of electrical stimulation (ES) of the parasympathetic nerves on the gustatory neural responses. The gustatory neural impulses of the GPNs were recorded using bipolar AgCl wires under normal blood circulation and integrated with a time constant of 1 s. Electrical stimuli were applied to the proximal side of the GPN with a pair of AgCl wires. The parasympathetic nerves of the GPN were strongly stimulated for 10 s with 6 V at 30 Hz before taste stimulation. The integrated neural responses to 0.5 M NaCl, 2.5 mM CaCl₂, water, and 1 M sucrose were enhanced to 130–140% of the controls. On the other hand, the responses for 1 mM Q-HCl and 0.3 mM acetic acid were not changed by the preceding applied ES. After hexamethonium (a blocker of nicotinic ACh receptor) was intravenously injected, ES of the parasympathetic nerve did not modulate the responses for all six taste stimuli. The mechanism for enhancement of the gustatory neural responses is discussed.     

Slow potentials in taste cells induced by frog glossopharyngeal nerve stimulation

Intracellular recordings were made from the taste cells of atropinized bullfrogs while the glossopharyngeal (GP) nerve fibres were electrically stimulated. Two types of slow potential, slow hyperpolarizing potentials (HPs) and slow depolarizing potentials (DPs), were induced in the taste cells. The slow HPs appeared when the lingual capillary blood flow was kept above 0.7 mm/s, whereas the slow DPs appeared when the blood flow was slowed down below 0.7 mm/s. The membrane resistance of a taste cell increased during the generation of a slow HP, but decreased during the generation of a slow DP. The reversal potentials for the slow HPs and the slow DPs were recorded at the same membrane potential (-11 to approximately -13 mV). Activation of non-selective cation channels possibly induced the slow DP and inactivation of those channels possibly induced the slow HP in the taste cell membrane. Electrical stimulation of the GP nerve activated a population of C fibres in the nerve and possibly released neurotransmitters from the nerve terminals. Released neurotransmitters might cause modulation of the membrane conductance in taste cells that leads to generation of the slow potentials. The present data suggest that slow HPs and slow DPs evoked in the taste cells of atropinized frogs by GP nerve stimulation are induced by putative neurotransmitters in the taste disc.     

Unique after-hyperpolarization accompanying action potential inChara globularis

A unique after-hyperpolarization was found in internodal cells ofChara globularis. The cells generated an ordinary action potential due to regenerative depolarization induced by the outward electric current pulse larger than a threshold stimulus. After reaching a depolarizing peak, the membrane potential repolarized and overshooted the resting potential to a value which was somehow 40 mV more negative than the resting potential before stimulation (after-hyperpolarization). Since the membrane resistance increased during the after-hyperpolarization, the after-hyperpolarization is thought to be caused by an increase in the resistance (decrease in the conductance) of the passive diffusion channel.