Search

We analyzed the prevalence of gustatory test abnormalities in breast cancer (BC) patients undergoing chemotherapy. We enrolled 43 BC patients undergoing chemotherapy and 38 BC patients who had never undergone chemotherapy (control group). Two gustatory tests were conducted: an instillation method examining the threshold for four basic taste stimuli and an electrogustometry method measuring the threshold for perception with electric stimulation at the front two-thirds of the tongue (cranial nerve VII) and at the back third of the tongue (cranial nerve IX). The results of the two gustatory tests and clinicopathological factors were compared between the chemotherapy and control groups and between patients with and without awareness of dysgeusia in the chemotherapy group. In the chemotherapy group, 19 (44%) patients were aware of dysgeusia and 8 (19%) had hypogeusia using the instillation method. Although more patients had parageusia in the chemotherapy than control group, no significant differences in the results of the two gustatory tests were observed. Patients with dysgeusia awareness had a higher threshold at cranial nerve IX using the electrogustometry method than those without dysgeusia awareness; no significant differences in hypogeusia were observed using the instillation method. In fact, 74% (14/19) of patients with dysgeusia awareness could identify the four tastes accurately using the instillation method. Similar results were observed for the instillation and electrogustometry methods at cranial nerve VII. While approximately half of the chemotherapy patients were aware of dysgeusia, 81% (35/43) of them could accurately identify the four basic tastes using the instillation method.

The effects of unsaturated fatty acids on membrane properties were studied using conventional whole-cell patch-clamp recording of isolated wing (Ib) cells in bullfrog (Lithobates catesbeianus) taste discs. Applying arachidonic acid to the bath induced monophasic inward currents in 60% of wing cells and biphasic inward and outward currents in the other cells. The intracellular dialysis of arachidonic acid did not induce an inward current; however, it enhanced a slowly developing Ba2+-sensitive outward current. The effects of various unsaturated fatty acids were explored under the condition of Cs+ internal solution. Linoleic and α-linolenic acids induced large inward currents. Oleic, eicosapentaenoic and docosahexaenoic acids elicited the same inward currents as those of arachidonic acid. Wing cells, under the basal condition with Cs+ internal solution, displayed a small inward current of -1.1±0.1pA/pF at -50mV (n=40), in which the peak existed at a membrane potential of -49mV. Removing external Ca2+ further increased the inward current by -2.9±0.3pA/pF at -50mV (n=4) from the basal current and the peak was located at -55mV. External linoleic acid (50μM) also induced a similar inward current of -5.6±0.6pA/pF at -50mV (n=19) from the basal current and the peak was located at -61mV. External Ca2+-free saline and linoleic acid induced similar current/voltage (I/V) relationships elicited by a ramp voltage as well as voltage steps. Linoleic acid-induced currents were not influenced by replacing internal EGTA with BAPTA, whereas inward currents disappeared under the elimination of external Na+ and addition of flufenamic acid. These results suggest that dietary unsaturated fatty acids may depolarize wing (Ib) cells, which affects the excitability of these cells., Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 166(3), pp.434-440; 2013

Thermal modulation of signaling pathways leads to excitation of taste receptor cells, but the underlying mechanisms remain unclear. Furthermore, it has long been known that there are contrast effects in various senses. In this study, we investigated cold-taste contrast and the relationship between taste and somatosensation. We lowered intraoral temperature using cold stimulus as a pretreatment, then returned to normal temperature in 249 healthy subjects, before administering room temperature taste-stimulating solutions to investigate changes in sensitivity to the four basic tastes (Sweet, Salt, Sour, and Bitter). Statistical comparisons of taste recognition thresholds before and after cold stimulus showed increased taste sensitivity for all four basic tastes. After categorizing different levels of pre-cold stimulus taste sensitivity into groups and comparing changes in sensitivity to the four basic tastes before and after stimulus, we found that the lower the sensitivity to the four basic tastes, the greater the increase in sensitivity induced by cold stimulus. These findings suggest that taste and low temperature send afferent signals which cause interaction in the afferent pathways between the peripheral and central nervous systems. Cold stimulus may offer one possible treatment strategy for dysgeusia.

SUMMARY The membrane properties of isolated frog parathyroid cells were studied using perforated and conventional whole-cell patch-clamp techniques. Frog parathyroid cells displayed transient inward currents in response to depolarizing pulses from a holding potential of –84 mV. We analyzed the biophysical properties of the inward currents. The inward currents disappeared by the replacement of external Na+ with NMDG+ and were reversibly inhibited by 3 μmol l–1 TTX, indicating that the currents occur through the TTX-sensitive voltage-gated Na+channels. Current density elicited by a voltage step from –84 mV to–24 mV was –80 pA pF–1 in perforated mode and–55 pA pF–1 in conventional mode. Current density was decreased to –12 pA pF–1 by internal GTPγS (0.5 mmol l–1), but not affected by internal GDPβS (1 mmol l-1). The voltage of half-maximum (V1/2)activation was –46 mV in both perforated and conventional modes. V1/2 of inactivation was –80 mV in perforated mode and –86 mV in conventional mode. Internal GTPγS (0.5 mmol l–1) shifted the V1/2 for activation to–36 mV and for inactivation to –98 mV. A putative endocannabinoid,2-arachidonoylglycerol ether (2-AG ether, 50 μmol l–1) and a cannabinomimetic aminoalkylindole, WIN 55,212-2 (10 μmol l–1) also greatly reduced the Na+ current and shifted the V1/2 for activation and inactivation. The results suggest that the Na+ currents in frog parathyroid cells can be modulated by cannabinoids via a G protein-dependent mechanism.

e21668 Background: Dysgeusia is a common side effect of chemotherapy (CT) and often impacts negatively on QOL. This study analyzed the prevalence of gustatory test abnormalities in breast cancer (BC) patients undergoing CT. Methods: We enrolled 33 BC patients who were undergoing CT (26 adjuvant CT and 7 CT for advanced BC) and conducted 2 gustatory tests on CT day. In the test of sensitivity using the instillation method (IM), 4 basic taste stimuli (sweet, salty, sour, and bitter) at 5 concentrations were applied to the entire oral cavity. For the electrogustometry method (EM), we measured the amount of current for perception of metallic tastes, examining the front two thirds of the tongue (cranial nerve VII) and the back third of the tongue (cranial nerve IX). We compared awareness of dysgeusia with results of the 2 gustatory tests. Results: Median CT duration was 83 days (12–1,353). Fifteen (45%) patients had awareness of dysgeusia (Grade 1: 11, Grade 2: 4). IM revealed 8 (24%) patients with abnormalities, 7 in a single taste species and 1 in 3 taste species. EM revealed 20 (61%) patients withabnormalities, 7 in the cranial nerve VII and 20 in the cranial nerve IX innervation area. All patients with abnormalities in the cranial nerve VII innervation area also had abnormalities in the cranial nerve IX innervation area. Among 15 patients with awareness of dysgeusia, there were 5 (33%) IM abnormalities and 12 (80%) EM abnormalities (4 in the cranial nerve VII and 12 in the cranial nerve IX innervation area). Among 18 patients without awareness of dysgeusia, there were 3 (16%) IM abnormalities and 8 (44%) EM abnormalities (3 in the cranial nerve VII and 8 in the cranial nerve IX nerve innervation area). Patients with IM abnormalities had a higher frequency of abnormalities in the cranial nerve VII innervation area than the normal range (50% and 12%, respectively; p = 0.04); there was no significant difference in the cranial nerve IX innervation area. Conclusions: Nearly half of the patients reporteddysgeusia, but most of these could correctly identify the four basic tastes. Further studies to clarify the discrepancy between subjective and objective observations could lead to effective prevention and treatment of dysgeusia.

We identified a Cl^- channel, two K^+ channels and a cAMP-gated channel which were isolated from bullfrog fungiform papilla cell membranes and incorporated into phospholipid bilayers using the tip-dip method. The 156 pS channels were inhibited by 100μM 4, 4'-diisothiocyanostilbene-2, 2'-disulfonic acid (DIDS) and displayed the reversal potential identical to the equilibrium potential of Cl^-, it was identified as a Cl^- channel. Two types of K^+ channel had unitary conductances of 79 and 43 pS, which may correspond to those of Ca^-activated and cAMP-blockable K^+ channels observed in isolated intact frog taste cell membranes, respectively. These results suggest that the tip-dip method is useful for stable investigation of the properties of ion channels already identified in the taste cell. Furthermore, the 23 pS channels were newly found and were activated directly by internal cAMP as cyclic nucleotide-gated (CNG) nonselective cation channels established in olfactory receptor cells. Thus, our results suggest the possibility that besides Cl^- and K^+ channels, the cAMP-gated channels contribute to taste transduction., Zoological science 18(3) pp.299-307; 2001

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.

The transduction mechanism of the conductance activated by saccharin was analysed in isolated bullfrog taste cells under whole-cell voltage-clamp. Bath application of 30 mM saccharin induced an inward current of -34 +/- 12 pA (mean +/- SEM, n = 10) at a membrane potential of -50 mV in 10 (23%) of 44 rod cells. The concentration-response relationship for the saccharin-gated current was consistent with that of the gustatory neural response. The saccharin-induced current was accompanied with a conductance increase under internal low Cl- condition (E(Cl) = -56 mV), suggesting that saccharin activated a cation conductance. The reversal potential of the saccharin-induced current was -17 +/- 2 mV (n = 10). Intracellular dialysis of 0.5 mM guanosine 5'-O-(2-thiodiphosphate) (GDP-beta-S) completely blocked the saccharin-induced response, suggesting the involvement of a G protein in the transduction. The dialysis of heparin (1 mg/mL) also inhibited the response almost completely, but the dialysis of 1 mM 8-Br-cAMP did not affect the response significantly. Intracellular 50 microM inositol 1,4,5-trisphosphate (1,4,5 InsP(3)) also induced the inward current in five (38%) of 13 rod cells, but intracellular Pasteurella multocida toxin (5 microg/mL, G alpha q-coupled PLC activator) did not elicit any response in the cells. The results suggest that saccharin mainly activates a cation conductance in frog taste cells through the mediation of IP3 production.

We have investigated the effect of 5-hydroxytryptamine (serotonin) (5-HT) on the membrane properties of bullfrog taste receptor cells (TRCs) using patch-clamp technique. External application of 5-HT reversibly suppressed the voltage-gated Na(+) current (I(Na)) in about half of the TRCs sampled. The magnitude of suppression of peak I(Na) was dependent on the holding potential of the cell. Forskolin and cyclic adenosine monophosphate (cAMP) mimicked the suppressive effect of 5-HT on I(Na), but an internal protein kinase A-inhibitor potentiated I(Na). These results suggest that 5-HT suppresses I(Na) of bullfrog TRCs via protein kinase A-dependent phosphorylation, resulting in suppression of the excitability of bullfrog TRCs.

When the glossopharyngeal (GP) nerve of the frog was stimulated electrically, electropositive slow potentials were recorded from the tongue surface and depolarizing slow potentials from taste cells in the fungiform papillae. The amplitude of the slow potentials was stimulus strength- and the frequency-dependent. Generation of the slow potentials was not related to antidromic activity of myelinated afferent fibers in the GP nerve, but to orthodromic activity of autonomic post-ganglionic C fibers in the GP nerve. Intravenous injection of atropine abolished the positive and depolarizing slow potentials evoked by GP nerve stimulation, suggesting that the slow potentials were induced by the activity of parasympathetic post-ganglionic fibers. The amplitude and polarity of the slow potentials depended on the concentration of adapting NaCl solutions applied to the tongue surface. These results suggest that the slow potentials recorded from the tongue surface and taste cells are due to the liquid junction potential generated between saliva secreted from the lingual glands by GP nerve stimulation and the adapting solution on the tongue surface.

The chorda tympani nerve responses to NaCl in a mouse strain, C57BL/6 are known to be much more sensitive than those in BALB/c. We compared the NaCl-induced responses obtained from taste cells of the fungiform papillae in these two strains of mice. Amiloride inhibited, in the same degree, the responses induced by a bath-application of normal extracellular solution (NES) containing 140 mM NaCl in either taste cells of C57BL/6 and BALB/c mice. In contrast, amiloride inhibited 62% of responses induced by an apically applied 0.5 M NaCl in the C57BL/6 strain, but only 33% of responses in the BALB/c strain. These results suggest that the difference in amiloride-sensitivity between taste cells in both strains mainly derives from the difference in density of functional amiloride sensitive Na+ channels at the apical receptive membrane but not at the basolateral membrane.

The amiloride-sensitive and -insensitive components of salt- and acid-induced responses in C57BL/6 and BALB/c mouse nondissociated taste cells were examined using the whole-cell clamp technique and localized taste stimulation method. Both amiloride-sensitive and -insensitive components were involved in a salt-induced depolarizing response. The taste cells of both mouse strains exhibited an amiloride-insensitive salt-induced response, which consisted of multiple components. In C57BL/6 mice, an amiloride-sensitive strain, more than 60% of salt-induced responses were amiloride-sensitive at the apical membrane, whereas in BALB/c mice, an amiloride-insensitive strain, less than 40% of responses were amiloride-sensitive. All the acid-induced responses in taste cells examined were insensitive to amiloride, and were markedly suppressed by the Cl channel blocker, 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB). These results suggest that multiple, different transduction mechanisms are involved in, and contribute to salty and sour transductions. In addition, we found that strain differences exist in salty transduction.

The effect of intracellular Ca2+ increase was analysed in isolated frog taste cells under the whole-cell patch clamp. External application of a Ca2+-ionophore, ionomycin (3 microM) induced the sustained inward current of -200+/-17 pA (mean +/- SE, n = 23) at -50 mV in taste cells. The ionomycin-induced response was observed in most of the cells exposed in the drug, but not when 10 mM BAPTA (1,2-bis (O-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid) was included in the pipette (eight cells). Steady-state I-V relationships of ionomycin-induced currents were almost linear and reversed at -8+/-1 mV (n = 23). The simultaneous removal of Na+ and Ca2+ from the external solution eliminated the response completely (three cells). Intracellular dialysis with 1 mM Ca2+ or 50 microM inositol 1,4,5-trisphosphate (IP3) in K+-internal solution also induced an inward current in the taste cells. The Ca2+-induced and IP3-induced responses were observed in 82% and 36% of the cells dialysed with the drugs, respectively. The Ca2+-induced and IP3-induced currents were inhibited by external Cd2+ (1-2 mM). The reversal potentials of the inward currents were -15+/-3 mV (n = 9) in Ca2+ dialysis and -11+/-3 mV (n = 13) in IP3 dialysis. The half-maximal Ca2+ concentration in the pipette to induce the inward current was approximately 170 microM. The results suggest that IP3 can depolarize the taste cell with mediation by intracellular Ca2+.

The percentage of non-tasters for phenylthiocarbamide in 915 Japanese students was 9.4%. The thresholds of the edge and back of the tongue to quinine hydrochloride were significantly smaller in the non-tasters than in the tasters. The thresholds of any tongue portions to NaCl, acetic acid or sucrose did not differ between the tasters and the non-tasters.

Na + -dependent K + conductance observed in apical membrane of isolated bullfrog taste cells was analyzed using various modifications of the patch clamp method. When the apical receptive membrane of a taste cell was exposed to 0.5 M NaCl using a pipette perfusion method, an inward current appeared in the cell-attached patch membrane. The permeability ratio of Na + to K + ( P Na / P K ) calculated from the reversal potential was 0.3. When an outside-out patch from the receptive membrane was exposed to 0.25 M NaCl solution, two types of K + channel, whose conductance were 35.8 and 9.4 pS respectively, were activated. These channels were reversibly activated by increasing internal Na + concentration. In the whole cell configuration, the outward current evoked by a voltage ramp from −808561 to 80 mV was significantly suppressed by replacement of Na + by N -methyl- d -glucamine^ (NMDG +) . The Na + -dependent outward current was blocked by 10 mM BaCl 2 , with positive shift of the zero-current potential in the whole cell recordings, indicating that the outward current including Na + -dependent K + component contributes to maintenance of the resting potential. These results suggest that small conductance Na + -dependent K + channels, which are also permeable to Na + , are involved in salt signal transduction in frog taste cells.

Response properties of nociceptors in the temporomandibular joint (TMJ) and surrounding area under experimental inflammation were investigated using an in vitro TMJ-nerve preparation in the rat. Nociceptive units (receptor and innervating nerve fiber) were classified into the following subtypes: Aδ-high-threshold mechanonociceptor (HTM), Aδ-polymodal nociceptor (POLY), C-HTM and C-POLY. In the inflamed joint, mechanical thresholds tended to be lower; however, the reaction to bradykinin was not identified as clearly as in control. Experimentally induced inflammation increased the proportion of heat-sensitive units and lowered heat threshold significantly. These results suggest that inflammation may sensitize nociceptors in the temporomandibular joint, and cause hyperalgesia and allodynia.

Dysgeusia causes a decrease in appetite, and it is one of the major factors in undernutrition. Dysgeusia is elicited by numerous causes, and in many cases it is still difficult to treat the various symptoms complained of by patients. We herein report a case in which dysgeusia was improved by transient cooling of the mouth.

The effect of calcium-sensing receptor (CaR) agonists on frog gustatory responses was studied using glossopharyngeal nerve recording and whole-cell patch-clamp recording of isolated taste disc cells. Calcimimetic NPS R-467 dissolved in normal saline solution elicited a large transient response in the nerve. The less active enantiomer of the compound, NPS S-467 induced only a small neural response. The EC(50) for NPS R-467 was about 20 microM. Cross-adaptation experiments were performed to examine the effect of 30 microM NPS R-467 and 100 microM quinine on the gustatory neural response. The magnitude of the R-467-induced response after adaptation to quinine was approximately equal to that after adaptation to normal saline solution, indicating that the receptor site for NPS R-467 is different from the site for quinine. NPS R-467 (100 microM) also induced an inward current accompanied with conductance increase and large depolarization in two (13%) of 15 rod cells, and a sustained decrease in outward current and small depolarization in six (40%) other rod cells. NPS S-467 (100 microM) induced a sustained decrease in outward current and depolarization in five (50%) of 10 rod cells. Another calcimimetic cinacalcet (100 microM) induced an inward current accompanied with conductance increase in three (27%) of 11 rod cells. The results suggest that NPS R-467 induces neural responses through cell responses unrelated to a resting K(+) conductance decrease.

Recent studies have indicated that nociceptors can be classified into various types according to their physiological properties. These studies have clarified that the frequency distribution of various nociceptor types is different among body sites and animal species. In the present study, we investigated the physiological properties of rat's periodontal nociceptors in an in vitro jaw-nerve preparation. Responses were recorded from functional single filaments in the inferior alveolar nerve. To determine the nociceptor type, calibrated von Frey filaments, heat, and bradykinin (BK) stimuli were used. We found five subtypes of nociceptors in the periodontal ligaments of the lower incisor: Adelta-high threshold mechanonociceptors (Adelta-HTM, n=28), Adelta-mechanoheat nociceptors (Adelta-MH, n=6), Adelta-polymodal nociceptors (Adelta-POLY, n=26), C-high threshold mechanonociceptors (C-HTM, n=3) and C-polymodal nociceptors (C-POLY, n=4). Most nociceptors were Adelta-innervated, while only a small number of C-innervated nociceptors were found. The present results suggest that periodontal nociceptors transmit mainly fast pain, and may thus play a role in rapid detection of injure-related stimuli during mastication.

Background and Objectives Analgesic properties of laser irradiation have been of great interest in the field of dentistry. This study aimed at evaluating the analgesic effects of the Er:YAG laser system in rats during and after laser irradiation. Study Design/Materials and Methods A pulsed Er:YAG laser was applied to the oral mucosa of the mandibular incisor at an energy density of approximately 0.1 J/cm2/pulse for 10 minutes at 10 Hz, and the integrated digastric muscle electromyogram in tooth pulp-evoked jaw-opening reflex was used as an index of the nociceptive response. Results Significant reflex suppression was observed 10 minutes after laser irradiation. The reflex amplitude started to return to its original level about 45 minutes after cessation of laser irradiation. Conclusions The Er:YAG laser used at low output levels presented inhibitory effects on the tooth pulp-evoked jaw-opening reflex, suggesting that this laser system may be of use for pain control during various dental treatments. Lasers Surg. Med. 33:169–172, 2003. © 2003 Wiley-Liss, Inc.

The transduction mechanism of the conductance activated by saccharin was analysed in isolated bullfrog taste cells under whole-cell voltage-clamp. Bath application of 30 mM saccharin induced an inward current of -34 +/- 12 pA (mean +/- SEM, n = 10) at a membrane potential of -50 mV in 10 (23%) of 44 rod cells. The concentration-response relationship for the saccharin-gated current was consistent with that of the gustatory neural response. The saccharin-induced current was accompanied with a conductance increase under internal low Cl- condition (E(Cl) = -56 mV), suggesting that saccharin activated a cation conductance. The reversal potential of the saccharin-induced current was -17 +/- 2 mV (n = 10). Intracellular dialysis of 0.5 mM guanosine 5'-O-(2-thiodiphosphate) (GDP-beta-S) completely blocked the saccharin-induced response, suggesting the involvement of a G protein in the transduction. The dialysis of heparin (1 mg/mL) also inhibited the response almost completely, but the dialysis of 1 mM 8-Br-cAMP did not affect the response significantly. Intracellular 50 microM inositol 1,4,5-trisphosphate (1,4,5 InsP(3)) also induced the inward current in five (38%) of 13 rod cells, but intracellular Pasteurella multocida toxin (5 microg/mL, G alpha q-coupled PLC activator) did not elicit any response in the cells. The results suggest that saccharin mainly activates a cation conductance in frog taste cells through the mediation of IP3 production.

The effect of a rise in intracellular Ca(2+) concentration was analyzed in isolated rat olfactory neurons using a whole-cell patch-clamp technique. Intracellular dialysis of 1 mmol l(-)(1) Ca(2+) in a standard-K(+), low-Cl(-) internal solution (E(Cl)=-69 mV) from the patch pipette into the olfactory neurons induced a sustained outward current of 49+/-5 pA (N=13) at -50 mV in all the cells examined. The outward currents were inhibited by external application of 100 micromol l(-)(1) 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB). External application of a Ca(2+) ionophore, 3 micromol l(-)(1) ionomycin, induced an inward current in three of eight cells whose voltages were clamped using the gramicidin-perforated technique, but ionomycin elicited an outward current in the other five cells, suggesting that natural intracellular Cl(-) concentration in the olfactory neurons was heterogeneous. While intracellular dialysis of 50 micromol l(-)(1) inositol 1,4,5-trisphosphate (1,4,5-InsP(3)) in the standard-K(+), low-Cl(-) internal solution induced the NPPB-sensitive outward current in 31 % of cells, and 500 micromol l(-)(1) cAMP induced it in 21 % of cells, a large proportion of the cells displayed an inward current in response to 1,4,5-InsP(3) and cAMP. The results suggest that 1,4,5-InsP(3) and cAMP can elicit Ca(2+)-dependent Cl(-) conductance and Ca(2+)-independent cation conductance in rat olfactory neurons.

Densities of pressure, pain and temperature spots in the back of the hand in 551 students and two-point discrimination thresholds in the hand, the face and the mouth in 684 students were measured. The mean numbers of pressure, pain, warm and cold spots in the back of the hand were 24.7/cm2, 130.5/cm2, 3.4/cm2 and 9.1/cm2, respectively. The mean thresholds of two-point discrimination were 1.7 mm in the tip of the tongue, 2.4 mm in the upper lip, 5.5 mm in the lower jaw, 7.5 mm in the palm, 8.8 mm in the forehead, and 11.8 mm in the back of the hand. There were mostly no differences between males and females in the values of sensory spots and two-point discrimination thresholds.

Miyamoto, Takenori, Rie Fujiyama, Yukio Okada, and Toshihide Sato. Sour transduction involves activation of NPPB-sensitive conductance in mouse taste cells. J. Neurophysiol. 80: 1852–1859, 1998. We examined the sour taste transduction mechanism in the mouse by applying whole cell patch-clamp technique to nondissociated taste cells from the fungiform papillae. Localized stimulation with 0.5 M NaCl and 25 mM citric acid (pH 3.0) of the apical membrane enabled us to obtain responses from single taste cells under a quasi-natural condition. Of 28 taste cells examined, 11 cells (39%) responded to 0.5 M NaCl alone and 2 cells (7%) responded to 25 mM citric acid alone, indicating the presence of salty- and sour-specific taste cells. Ten cells (36%) responded to both NaCl and citric acid and 5 cells (18%) responded to neither salt nor citric acid. Amiloride reversibly suppressed NaCl-induced responses in mouse taste cells but not citric acid-induced responses. On the other hand, a Cl−channel blocker, 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB), reversibly suppressed all the citric-acid-induced responses. Most of the NaCl-induced current responses displayed an inwardly rectifying property, whereas all the citric-acid-induced responses displayed an outwardly rectifying property. The reversal potential for NPPB-sensitive component in citric-acid-induced current responses was −2 ± 7 mV (mean ± SE, n = 4), which was close to the equilibrium potential of Cl−( ECl), whereas the reversal potential for NPPB-insensitive component was 34 ± 8 mV ( n = 4). The reversal potential of citric-acid-induced current responses (19 ± 8 mV, n = 4) was mostly present at the middle point between reversal potentials of NPPB-sensitive and -insensitive current components. In some taste cells, an inorganic cation channel blocker, Cd2+, suppressed citric-acid-induced responses, but an inorganic stretch-activated cation channel blocker, Gd3+, did not affect these responses. These results suggest that salt- and acid-induced responses were mediated by differential transduction mechanisms in mouse taste cells and that NPPB-sensitive Cl−channels play a more important role to sour taste transduction rather than amiloride-sensitive Na+channels. However, the fact that the reversal potentials of citric-acid-induced responses had more positive than EClsuggests that Ca2+or H+permeable and poorly selective cation channels, which should be amiloride insensitive, may be activated by citric acid.

The effect of arginine vasopressin (AVP) on the membrane properties was analyzed in isolated bullfrog taste cells using a perforated whole-cell patch-clamp technique. AVP (100 nM) induced three kinds of responses in rod-type taste cells: appearance of inward current, inhibition of voltage ramp-induced outward current and enhancement of the outward current. The Ca(2+)-ionophore ionomycin (3 microM) also induced inward current in taste cells. A membrane-permeable cAMP analog, 8-CPT-cAMP (0.3 mM) inhibited voltage ramp-induced outward current in some rod cells, but enhanced the current in other rod cells. The results suggest that AVP may increase either intracellular Ca2+ level or cAMP level in taste cells, modulating the membrane excitability.

We could identify two types of K+ channels, of 80 and 40 pS conductance, respectively, in the bullfrog taste cell membrane using excised and cell-attached configurations of the patch-clamp technique. The taste cell membrane could be divided into four membrane parts--receptive area, apical process, cell body and proximal process. The 80-pS K+ channels were dependent on voltage and Ca2+ and were located exclusively on the receptive membrane and the apical process membrane. The 40-pS K+ channels were independent of voltage and Ca2+. The open probability of 40-pS K+ channels was decreased by the simultaneous presence of cyclic adenosine monophosphate (cAMP) and adenosine triphosphate (ATP), and the suppressive effect was antagonized by protein kinase inhibitor (PKI). Although 40-pS K+ channels were found in a high density on the receptive and apical process membranes, the channels also were present in the other two parts of the taste cell membrane. These results suggest that the two different types of K+ channel in the bullfrog taste cells may play different roles in gustatory transduction.

The contribution of the transduction pathway on the basolateral [1] and the receptive membranes [2] to the mechanism underlying the generation of the receptor potential induced by salt stimuli has been suggested individually by different groups. In previous studies, we reexamined the transduction mechanism of salt stimuli by changing the superficial and interstitial fluid of the frog tongue separately, using the lingual artery perfusion method [3, 4]. Three important results were obtained: (1) the magnitude of the receptor potentials decreased by 50% after the removal of Na+ and Ca2+ from interstitial fluid; (2) the reversal potential of salt-induced responses altered depending on the types and concentrations of salt stimuli after removal of Na+ and Ca2+ from both the superficial and interstitial fluid; and (3) substitution of an organic anion for Cl− and application of a Cl− channel blocker changed the reversal potential of salt response. These results led us to the conclusion that at least three types of ion channels, non-selective cation channels, K+ channels, and Cl− channels, on the receptive membrane, and a Na+ conductive pathway activated by an unknown second messenger on the basolateral membrane contribute to salt-induced responses in frog taste cells.

Elevation of extracellular Ca(2+) concentration induces intracellular Ca(2+) signaling in parathyroid cells. The response is due to stimulation of the phospholipase C/Ca(2+) pathways, but the direct mechanism responsible for the rise of intracellular Ca(2+) concentration has remained elusive. Here, we describe the electrophysiological property associated with intracellular Ca(2+) signaling in frog parathyroid cells and show that Ca(2+)-activated Cl(-) channels are activated by intracellular Ca(2+) increase through an inositol 1,4,5-trisphophate (IP(3))-independent pathway. High extracellular Ca(2+) induced an outwardly-rectifying conductance in a dose-dependent manner (EC(50) ∼6 mM). The conductance was composed of an instantaneous time-independent component and a slowly activating time-dependent component and displayed a deactivating inward tail current. Extracellular Ca(2+)-induced and Ca(2+) dialysis-induced currents reversed at the equilibrium potential of Cl(-) and were inhibited by niflumic acid (a specific blocker of Ca(2+)-activated Cl(-) channel). Gramicidin-perforated whole-cell recording displayed the shift of the reversal potential in extracellular Ca(2+)-induced current, suggesting the change of intracellular Cl(-) concentration in a few minutes. Extracellular Ca(2+)-induced currents displayed a moderate dependency on guanosine triphosphate (GTP). All blockers for phospholipase C, diacylglycerol (DAG) lipase, monoacylglycerol (MAG) lipase and lipoxygenase inhibited extracellular Ca(2+)-induced current. IP(3) dialysis failed to induce conductance increase, but 2-arachidonoylglycerol (2-AG), arachidonic acid and 12S-hydroperoxy-5Z,8Z,10E,14Z-eicosatetraenoic acid (12(S)-HPETE) dialysis increased the conductance identical to extracellular Ca(2+)-induced conductance. These results indicate that high extracellular Ca(2+) raises intracellular Ca(2+) concentration through the DAG lipase/lipoxygenase pathway, resulting in the activation of Cl(-) conductance., PLoS One, 6(4), e19158; 2011

Using single channel recordings of the patch clamp method, we found first that cation channels were present in the bullfrog taste cell membranes. These channels were widely distributed over the taste cell membrane. The conductance examined with inside-out patches was 30.7 +/- 5.7 pS (mean +/- S.D., n = 13). The permeability ratio of the cation channel to monovalent cations was PK:PCS:PNa = 1.18:1.00:0.70, indicating that the channels are non-selective.

We examined changes of intracellular free calcium concentration [Ca2+]i elicited by taste stimuli of sucrose, denatonium and NaCl in the taste buds of seven human fungiform papillae. In one taste bud we observed an increase in [Ca2+]i induced by only NaCl. In another bud an increase of [Ca2+]i in response to both NaCl and sucrose was found. The Ca2+ responses to NaCl and sucrose occurred in differential areas within the one taste bud. In the other five fungiform papillae [Ca2+]i was not changed by the taste stimuli. These results suggest that an increase of [Ca2+]i participates in taste transduction mechanisms for sucrose and NaCl, and that taste cells in one taste bud may respond to differential stimuli.

The transduction mechanism of salt-induced responses of mouse taste cells was investigated using the patch clamp and the local stimulation techniques under quasi-natural conditions. Apically applied NaCl induced a voltage-independent current, which was partially suppressed by amiloride and Cd 2+ . In contrast, apically applied 0.5 M KCl induced an inwardly rectifying current (KCl-induced Iir). The KCl-induced Iir was unaffected by amiloride. The Iir was suppressed not only by external Ba 2+ and Cs + , but also by a Cl – channel blocker, niflumic acid. The Er of the KCl-induced response was independent of the apical ionic concentration, but rather was close to the equilibrium potential of Cl – (ECl) at the basolateral membrane. The KCl-induced Iir displayed a fast run-down under the conditions of the conventional whole cell clamp method, but not under the perforated patch conditions. Immunohistochemical localization of an inwardly rectifying Cl – channel protein, ClC-2, was observed in taste bud cells of the fungiform papillae. It is concluded that the transduction mechanism of NaCl-induced responses is completely different from that of KCl-induced responses in mouse taste cells.