Transcriptional activities of two newly identified Haemaphysalis longicornis tick-derived promoter regions in the Ixodes scapularis tick cell line (ISE6).

Ticks are obligate haematophagous ectoparasites considered to be second to mosquitoes as vectors of human diseases and the most important vector for animals. Despite efforts to control tick infestations, they remain a serious health problem. Gene manipulation has been established in mosquitoes and led to the control of mosquito populations and of mosquito-borne pathogens. Therefore, gene manipulation could be useful for controlling ticks and tick-borne pathogens. To investigate effective gene expression vectors for ticks, the promoter activities of commercial plasmids were evaluated in a tick cell line (ISE6). Dual luciferase assays revealed that pmirGLO, the human phosphoglycerate kinase promoter contained plasmid vector, showed the highest activity in ISE6 cells amongst the tested plasmids. Moreover, we identified the promoter regions of the Haemaphysalis longicornis actin (HlAct) and the intracellular ferritin (HlFer1) genes. To construct a more effective expression vector for ticks, these promoter regions were inserted into pmirGLO (pmirGLO-HlAct pro and pmirGLO-HlFer1 pro). The pmirGLO-HlAct pro vector showed significantly higher promoter activity than pmirGLO, whereas the pmirGLO-HlFer1 pro vector demonstrated significantly lower promoter activity than pmirGLO in ISE6 cells. The HlAct promoter region may have high promoter activity in ISE6 cells. The results of the present study provide useful information for the development of a genetic modification system in ticks.

Susceptibility of isogeneic ginbuna Carassius auratus langsdorfii Temminck et Schlegel to cyprinid herpesvirus-2 (CyHV-2) as a model species.

Herpesviral haematopoietic necrosis (HVHN), caused by cyprinid herpesvirus-2 (CyHV-2), has affected the commercial production of the goldfish Carassius auratus and gibelio carp Carassius auratus gibelio. High water temperature treatments are reported to reduce the mortality rate of infected goldfish and elicit immunity in the survivors. To define the mechanism by which this intervention induces resistance, clonal ginbuna Carassius auratus langsdorfii, which is closely related to both species and has been used in fish immunology, may represent a promising model species. In this study, we investigated the susceptibility of clonal ginbuna strains to CyHV-2 and the effect of high water temperature treatment on infected ginbuna and goldfish. Experimental intraperitoneal infection with CyHV-2 at 25 °C caused 100% mortality in ginbuna strains, which was accompanied by histopathological changes typical of HVHN. Both infected ginbuna S3n strain and goldfish, exposed to high temperature for 6 days [shifting from 25 °C (permissive) to 34 °C (non-permissive)], showed reduced mortalities after the 1st inoculation, and subsequent 2nd virus challenge to 0%, indicating induction of immunity. It was concluded that ginbuna showed a similar susceptibility and disease development in CyHV-2 infection compared to goldfish, suggesting that ginbuna can be a useful fish model for the study of CyHV-2 infection and immunity.

Electronic ferroelectricity from charge ordering in RFe(2)O(4).

We report our recent discovery of novel ferroelectricity arising from the polar ordering of Fe(3)+ and Fe(2)+ in a mixed valence triangular lattice oxide LuFe(2)O(4), where the electric polarization is not a result of ionic displacement. The polar ordering of Fe(3)+ and Fe(2)+ was confirmed with a resonant x-ray scattering study in SPring-8. The origin of such ordering is the competitive interaction between Fe(3)+ and Fe(2)+ in the triangular lattice, i.e., the charge frustration. The polar superlattice of Fe(3)+ and Fe(2)+ develops below 350 K, where the electric polarization appears. The ferroelectricity arising from the polar charge ordering or the polar electron distribution may have great potential for the future application of ferroelectrics.

Selective enhancement and suppression of frog gustatory responses to amino acids.

Properties of the receptor sites for L-amino acids in taste cells of the bullfrog (Rana catesbeiana) were examined by measuring the neural activities of the glossopharyngeal nerve under various conditions. (a) The frogs responded to 12 amino acids, but the responses to the amino acids varied with individual frogs under natural conditions. The frog tongues, however, exhibited similar responses after an alkaline treatment that removes Ca2+ from the tissue. The variation in the responses under natural conditions was apparently due to the variation in the amount of Ca2+ bound to the receptor membrane. (b) The responses to hydrophilic L-amino acids (glycine, L-alanine, L-serine, L-threonine, L-cysteine, and L-proline) were of a tonic type, but those to hydrophobic L-amino acids (L-valine, L-leucine, L-isoleucine, L-methionine, L-phenylalanine, and L-tyrptophan) were usually composed of both phasic and tonic components. (c) The properties of the tonic component were quite different from those of the phasic component: the tonic component was largely enhanced by the alkaline treatment and suppressed by the acidic treatment that increases binding of Ca2+ to the tissue. Also, the tonic component was suppressed by the presence of low concentrations of salts, or the action of pronase E, whereas the phasic component was unchanged under these conditions. These properties of the phasic component were quite similar to those of the response to hydrophobic substances such as quinine. These results suggest that the hydrophilic L-amino acids stimulate receptor protein(s) and that the hydrophobic L-amino acids stimulate both the receptor protein and a receptor site similar to that for quinine. (d) On the basis of the suppression of the responses to amino acids by salts, the mechanism of generation of the receptor potential is discussed.

Gustatory responses of eel palatine receptors to amino acids and carboxylic acids.

The gustatory receptors of the eel palate were found to be extremely sensitive to amino acids and carboxylic acids. The results obtained are as follows: (a) 11 amino acids which are among naturally occurring amino acids elicited responses in the palatine nerve, but 9 amino acids did not elicit a response even at a high concentration. The effect of D-amino acids was always much less than that of their corresponding L-isomers. There was no appreciable difference in the effectiveness of an alpha-amino acid (alpha-alanine) and beta-amino acid (beta-alanine). (b) The threshold concentrations of the most potent amino acids (arginine, glycine) were between 10(-8) and 10(-9) M. A linear relation between the magnitude of the response and log stimulus concentration held for a wide concentration range for all the amino acids examined. (c) The palatine receptors responded sensitively to various carboxylic acid solutions whose pH was adjusted to neutral. The threshold concentrations varied between 10(-4) and 10(-7) M. The magnitude of the response at 10(-2) M increased with an increase of carbon chain length. (d) The extent of cross-adaptation was examined with various combinations of amino acids. A variety of the response patterns showing complete cross-adaptation, no cross-adaptation, or synergetic interaction was observed. The synergetic interaction was also observed when one amino acid below its threshold concentration was added to the other amino acid below its threshold concentration was added to the other amino acid. No cross-adaptation was observed between amino acids and fatty acids. (e) The treatment of the palate with papain led to loss of the responses to arginine, glycine, and histidine without affecting those to proline and acetic acid. The treatment with pronase E eliminated selectively the response to proline. The possibility that the eel gustatory receptors are responsible for sensing food at a distance was discussed.

Taste transduction mechanism: similar effects of various modifications of gustatory receptors on neural responses to chemical and electrical stimulation in the frog.

Responses in the frog glossopharyngeal nerve induced by electrical stimulation of the tongue were compared with those induced by chemical stimuli under various conditions. (a) Anodal stimulation induced much larger responses than cathodal stimulation, and anodal stimulation of the tongue adapted to 5 mM MgCl2 produced much larger responses than stimulation with the tongue adapted to 10 mM NaCl at equal current intensities, as chemical stimulation with MgCl2 produced much larger responses than stimulation with NaCl at equal concentration. (b) The enhansive and suppressive effects of 8-anilino-1-naphthalenesulfonate, NiCl2, and uranyl acetate on the responses to anodal current were similar to those on the responses to chemical stimulation. (c) Anodal stimulation of the tongue adapted to 50 mM CaCl2 resulted in a large response, whereas application of 1 M CaCl2 to the tongue adapted to 50 mM CaCl2 produced only a small response. This, together with theoretical considerations, suggested that the accumulation of salts on the tongue surface is not the cause of the generation of the response to anodal current. (d) Cathodal current suppressed the responses induced by 1 mM CaCl2, 0.3 M ethanol, and distilled water. (e) The addition of EGTA or Ca-channel blockers (CdCl2 and verapamil) to the perfusing solution of the lingual artery reversibly suppressed both the responses to chemical stimulus (NaCl) and to anodal current with 10 mM NaCl. (f) We assume from the results obtained that electrical current from the microvillus membrane of a taste cell to the synaptic area supplied by anodal stimulation or induced by chemical stimulation activates the voltage-dependent Ca channel at the synaptic area.

Equilibrium properties of mouse-Torpedo acetylcholine receptor hybrids expressed in Xenopus oocytes.

This study used messenger RNA encoding each subunit (alpha, beta, gamma and delta) of the nicotinic acetylcholine (ACh) receptor from mouse BC3H-1 cells and from Torpedo electric organ. The mRNA was synthesized in vitro by transcription with SP6 polymerase from cDNA clones. All 16 possible combinations that include one mRNA for each of alpha, beta, gamma, and delta were injected into oocytes. After allowing 2-3 d for translation and assembly, we assayed each oocyte for (a) receptor assembly, measured by the binding of [125I]alpha-bungarotoxin to the oocyte surface, and (b) ACh-induced conductance, measured under voltage clamp at various membrane potentials. All combinations yielded detectable assembly (30-fold range among different combinations) and ACh-induced conductances (greater than 1,000-fold range at 1 microM). On double-logarithmic coordinates, the dose-response relations all had a slope near 2 for low concentrations of ACh. Data were corrected for variations in efficiency of translation among identically injected oocytes by expressing ACh-induced conductance per femtomole of alpha-bungarotoxin-binding sites. Five combinations were tested for d-tubocurarine inhibition by the dose-ratio method; the apparent dissociation constant ranged from 0.08 to 0.27 microM. Matched responses and geometric means are used for describing the effects of changing a particular subunit (mouse vs. Torpedo) while maintaining the identity of the other subunits. A dramatic subunit-specific effect is that of the beta subunit on voltage sensitivity of the response: gACh(-90 mV)/gACh(+30 mV) is always at least 1, but this ratio increases by an average of 3.5-fold if beta M replaces beta T. Also, combinations including gamma T or delta M usually produce greater receptor assembly than combinations including the homologous subunit from the other species. Finally, EACh is defined as the concentration of ACh inducing 1 microS/fmol at -60 mV; EACh is consistently lower for alpha M. We conclude that receptor assembly, voltage sensitivity, and EACh are governed by different properties.

Synthesis and biological activity of conformationally restricted analogues of milnacipran: (1S, 2R)-1-phenyl-2-[(R)-1-amino-2-propynyl]-N,N- diethylcyclopropanecarboxamide is a novel class of NMDA receptor channel blocker.

Conformationally restricted analogues of (+/-)-(Z)-2-aminomethyl-1-phenyl-N,N-diethylcyclopropanecarboxamide++ + [milnacipran, (+/-)-1] were designed on the basis of its characteristic cyclopropane structure and were synthesized enantioselectively to develop efficient NMDA receptor antagonists. Among these analogues, (1S,2R)-1-phenyl-2-[(R)-1-amino-2-propynyl]-N, N-diethylcyclopropanecarboxamide (2d) had one of the most potent affinities for the receptor, with a Ki value of 0.29 microM. The blockade of NMDA receptor channels expressed by Xenopus oocytes by 2d was investigated in detail, and 2d was identified as a new class of open channel blocker against this receptor.

Synthesis and biological activity of conformationally restricted analogs of milnacipran: (1S,2R)-1-phenyl-2-[(S)-1-aminopropyl]-N,N-diethylcyclopropanecarboxami de, an efficient noncompetitive N-methyl-D-aspartic acid receptor antagonist.

We recently demonstrated that (+/-)-(Z)-2-(aminomethyl)-1-phenyl-N,N-diethylcyclopropanecarboxamide [milnacipran, (+/-)-1], an inhibitor of the reuptake of serotonin (5-HT), was a noncompetitive NMDA receptor antagonist. On the basis of the cyclopropane structure of (+/-)-1, conformationally restricted analogs with different stereochemistries, namely 1-phenyl-2-(1-aminoalkyl)-N,N-diethylcyclopropanecarboxamindes (2, 3, ent-2, and ent-3), were designed and synthesized. Among these analogs, 2a, 2b, and 2f, with (1S,2R,1'S)-configuration, were more efficient than milnacipran as NMDA receptor antagonists; these compounds significantly inhibited the binding of [3H]MK-801 at IC50 = 0.35 +/- 0.08, 0.20 +/- 0.024, and 0.16 +/- 0.02 microM, respectively, and blocked the response of voltage-clamped oocytes to NMDA, surpassing the effects of (+/-)-1. Although both the 1'-methyl analog 2a and the 1'-vinyl analog 2f, like (+/-)-1, strongly inhibited 5-HT uptake in vitro, the corresponding 1'-ethyl analog 2b was devoid of the inhibitory effect on 5-HT uptake, while it was about 30 times more potent as an NMDA receptor antagonist than (+/-)-1.

Primary repair of interrupted aortic arch and severe aortic stenosis in neonates.

Two infants, aged 36 days old (Case 1) and 18 days old (Case 2) with interrupted aortic arch types B and A, respectively, and with severe aortic stenosis, were successfully operated on by use of pulsatile cardiopulmonary bypass. The great arteries were normally related in Case 1 and were transposed in Case 2. Repair involved the following procedure: ligation of the patent ductus arteriosus, restoration of aortic continuity with an 8 mm polytetrafluoroethylene graft, placement of an internal patch to tunnel all left ventricular blood from the left ventricle through the ventricular septal defect into the pulmonary artery in Case 1 and patch closure of the ventricular septal defect in Case 2, transection of the main pulmonary artery, anastomosis between the proximal pulmonary artery and the ascending aorta, and interposition of a valved conduit between the right ventricle and the distal pulmonary artery. The operative field could be approached easily through a median sternotomy. Postoperative cardiac catheterization revealed satisfactory anatomical and hemodynamic results in both cases.

Inhibitory effect of a new antibiotic, guanine 7-N-oxide, on the replication of several RNA viruses.

Guanine 7-N-oxide (G-7-Ox) was examined for its antiviral activity against 9 viruses based on plaque reduction, neuraminidase activity reduction, a fluorescent antibody technique or ELISA. The following viruses were included in the tests: influenza, Sendai, simian virus 5 (SV5), respiratory syncytial, western equine encephalitis, Japanese encephalitis, vesicular stomatitis, rabies and polio. G-7-Ox showed broad anti-RNA viral activity against all viruses tested, except for poliovirus. Inhibition of persistent SV5 infection by G-7-Ox indicates that its antiviral activity is independent of cytotoxicity.

Circulation of type 1 wild poliovirus in northern Vietnam during 1991-1994.

From 1991 to 1994, 143 polioviruses were isolated from patients with acute flaccid paralysis in northern Vietnam. Of these 143 isolates, 133 were type 1 and five of each were type 2 and type 3. These isolates were intratypically differentiated by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Of the 133 type 1 isolates, 113 were wild strains and only 20 isolates were of Sabin vaccine-like strains. These 113 isolates were divided into seven groups by PCR-RFLP patterns and were also classified into three genomic groups by nucleotide sequencing and phylogenetic analysis. One of the three genomic groups accounted for the majority (75) of these isolates. The isolates belonging to the predominant group were found during 1991-1993. The second group, which included 17 isolates, were detected only in 1993. These isolates had genome sequence similar to isolates in southern Vietnam in the same year. The third group of isolates were detected only in 1991 and were considered to be Mahoney-like wild strains. All isolates examined were different from those obtained in other countries. In 1994, however, no wild-type polioviruses were isolated in our study. These results reveal that three unique strains were circulating in northern Vietnam in recent years, and indicate that the incidence of poliomyelitis due to wild poliovirus is decreasing.

A method for in-situ tight-seal recordings from single taste bud cells of mice.

In order to investigate taste transduction mechanisms, we developed a method to irrigate the receptor and basolateral membranes of mouse taste bud cells with different solutions under tight-seal recording conditions. A peeled tongue epithelium with taste bud cells was mounted on a recording platform designed to separate irrigating solutions for each membrane. The mucosal surface (receptor membrane side) of the peeled epithelium facing an inner chamber was always irrigated with deionized water or stimulating solutions, and the serosal surface (basolateral membrane side) was irrigated with a physiological saline solution. A recording electrode was placed on the basolateral membrane of a taste bud cell under an upright-microscope with a x 40-water-immersed objective. Investigated taste bud cells generated action potentials, and 1 M glucose, 200 mM NaCl, and 10 mM quinine elicited inward current. Irrigation with deionized water for more than 1 h had no effect. The resistance of the peeled tongue epithelium was 1570 +/- 343 omega cm2. These results show that the peeled tongue epithelium protects basolateral membranes from deionized water or stimulating solutions as the tongue epithelium does in situ and that this method is suitable to investigate the role of each membrane in taste transduction.

NH3- and CO2-induced suppression of taste nerve responses in clawed toads and eels.

We investigated the effects of intracellular pH values (pHi) on taste nerve responses of clawed toads and eels. (1) CO2, NH3 or trimethylamine reversibly suppressed the taste nerve responses of clawed toads to various amino acids, CaCl2 and caffeine. IC50 values of the suppression of the responses to 0.1 mM L-proline were as follows: approximately 10 mM for CO2, approximately 0.3 mM for NH3, approximately 0.2 mM for trimethylamine. (2) Cross-adaptation experiments showed that L-proline, caffeine and CaCl2 stimulated different receptor sites from each other, indicating the suppressive effect was non-specific. (3) Although CO2, NH3 or trimethylamine yielded the charged molecules, HCO3-, CO3(2-), NH4+ or trimethylammonium on hydration, none of these charged species suppressed taste responses. (4) NH3 increased the threshold concentration of L-proline by e-fold per 0.37 mM NH3 and decreased the maximum response to L-proline with increasing NH3 concentration. (5) The taste nerve responses of eels to 0.1 mM L-arginine, a potent stimulus on eel taste receptors, were similarly suppressed by NH3 with an IC50 value of approximately 0.3 mM. (6) These results indicated that these uncharged species changed pHi, which suppressed the taste responses. CO2-induced acidosis and NH3- or trimethylamine-induced alkalosis are likely to inhibit activities of ion channels or enzymes involved in taste transduction mechanisms.

Taste responses of bullfrog to pungent stimuli.

Taste responses of bullfrogs to various pungent compounds and taste substances were electrophysiologically recorded from glossopharyngeal nerves. The threshold concentrations were approximately 10(-7) M for piperine, approximately 10(-6) M for capsaicin and approximately 10(-4) M for allyl isothiocyanate. At any concentration examined, piperine was more potent than capsaicin. Both piperine and capsaicin elicited desensitizing responses, but the taste receptors recovered from the desensitization within 10 min after washing with deionized water. Cross-adaptation experiments revealed that capsaicin only partially desensitizes receptors for piperine, L-leucine, HCl or quinine. Perfusion of the lingual artery with a solution containing no added Ca decreased the responses to capsaicin. Such a solution has been shown to suppress the taste nerve responses by blocking synaptic transmissions between taste cells and taste nerves [8]. These results suggest that the gustatory effects of capsaicin are different from its pharmacological effects on sensory neurons. It is likely that capsaicin and other pungent compounds, when they act as seasonings, stimulate taste cells rather than the free nerve endings of the sensory neurons.

In situ tight-seal recordings of taste substance-elicited action currents and voltage-gated Ba currents from single taste bud cells in the peeled epithelium of mouse tongue.

We investigated the taste responses of single taste-bud cells (TBCs) in mice by applying stimuli only on receptor membranes acclimated to deionized water under tight-seal cell-attached voltage-clamp conditions, while their basolateral membranes were irrigated with a physiological saline solution. For this irrigation, we developed a new method: a peeled-tongue epithelium with TBCs mounted on a recording chamber where the peeled epithelium separated the irrigating solutions for each membrane as it separated in situ. Although no quinine-elicited action potentials had been reported, TBCs elicited a long-lasting train of biphasic currents derived from the action potentials in response to 10 mM quinine, in addition to responses to 10 mM HCl, or 200 mM NaCl dissolved in deionized water. These results indicate that quinine as well as HCl and NaCl depolarizes TBCs and generate action potentials. Under whole-cell recording conditions, TBCs generated action potentials, and voltage-gated currents such as LVA and HVA Ca currents, TTX-sensitive Na currents, and TEA/4-AP-sensitive K currents on depolarization. These voltage-gated channels were shown to exist predominantly on the basolateral membranes. We discussed the receptor mechanisms and the role of taste substance-elicited action potentials.

Mechanism of the water response in carp gustatory receptors: independent generation of the water response from the salt response.

The carp gustatory responses to various salts and those to distilled water after adaptation of the receptors to the salts were recorded from the palatine nerve under varying conditions. (a) As far as Na4Fe(CN)6 solution prepared freshly was used, any peak in the dose-response curve was not observed, while the solution stored overnight induced a large peak response at the low concentration region as Konishi reported. The magnitude of the water response after adaptation to the stored solution was practically equal to that after adaptation to the fresh solution, suggesting that the receptor site for the water response is different from that for the dilute salts. (b) The responses to salts depended largely on the species of both cations and anions of the salts. The responses were deceased with an increase in the lyotropic number of the anions and increased with an increase in the number above 11. The response to distilled water was practically independent of the species of both monovalent cations and anions of the salts used for the adaptation. The pH dependence of the response to 10 mM NaCl was largely different from that to distilled water after adaptation to 10 mM NaCl. (c) The water response was suppressed by the presence of electrolytes in stimulating solution; the data obtained with different species of salts were described by a single curve as a function of the ionic strength. (d) The mechanism to explain how distilled water leads to depolarization of the taste cell was discussed in terms of the electric double-layer potential.

Ion dependence of the eel taste response to amino acids.

The effects of changed ionic environments on the taste responses to amino acids were examined by recording the activity of the palatine nerve. The responses to glycine, L-proline and L-arginine dissolved in deionized water were greatly reduced after ions were removed from the surface of the palatine epithelium by treatment with 5 mM EDTA. The addition of various species of salts to amino acid solutions caused reversible recovery of the responses. The response to 0.1 mM glycine increased with an increase in salt concentration in amino acid solutions and reached saturation level. The effects of salts of divalent cations (CaCl2 and MgCl2) appeared at much lower concentration (10(-7) M) than those of salts of monovalent cations (NaCl and KCl) (10(-4) M), suggesting that cations support the taste response. All the organic cations examined, including those of large molecular weights (choline, Tris, bis-Tris, bis-Tris propane, tetraethylammonium, triethanolamine and D-glucosamine), also supported the taste responses. The results obtained led to a conclusion that not a specific cation but various species of cations can support the eel taste responses. It is suggested that the cations do not act as current carrying ions to depolarize the taste cells but that the binding of the cations to the receptor membrane plays an essential role in taste reception.

Role of cations in olfactory reception.

The effects of changed ionic environments on the olfactory responses in the carp, the rainbow trout, and the bullfrog were examined by recording the stimulant-induced waves from the olfactory bulb. (a) Application of stimulants (various species of amino acids and beta-ionone) dissolved in deionized water to the EDTA-treated olfactory epithelium of the carp did not induce any response. The addition of various species of salts to the stimulating solution reversibly restored the response. (b) The responses of the carp to L-alanine in the presence of MgCl2 and MgSO4 and those in the presence of KCl and K4Fe(CN)6 are described by single curves, respectively, as a function of concentration of the cations, suggesting that the cations support olfactory reception. (c) All the inorganic cations examined (Li+, NH3+, K+, Ca2+, Mg2+, Co2+, Mn2+, Cd2+) and organic cations (Tris+, choline+, bis-Tris propane2+) were effective to support the response of the carp, whereas TEA+, TBA+, triethanolamine+, and bis-Tris+ were ineffective. (d) The olfactory responses of the rainbow trout and the bullfrog were also reduced by removal of ions from the surface of the epithelia and recovered by addition of ions. (e) It is suggested that the cations do not act as current carriers across the apical olfactory cell membrane for the generation of the receptor potential.

Antagonism between the suppressive effects of NH3 and CO2 on bullfrog taste nerve responses to quinine.

The suppression mechanism of NH3 and CO2 on bullfrog taste nerve responses to 0.1 mM quinine was investigated by applying them directly on the tongue surface (surface application) or at the tongue interstices through the lingual artery (interstitial application). The surface application of NH3 and CO2 reversibly suppressed the taste nerve responses with IC50 values of 0.37 mM and 2.2 mM, respectively, whereas their hydrates were ineffective. The interstitial application of NH3 reversibly suppressed the taste nerve responses. The surface application of CO2 recovered the suppressed responses to quinine. The 4 s application of CO2 prior to that of 0.1 mM quinine had the maximum effect. These results show that NH3-induced alkalosis and CO2-induced acidosis of taste cells, taste nerve endings, or tongue interstices surrounding them suppressed the taste nerve responses, and that the neutralization of their intracellular pH recovered these responses. The time-dependent recovery suggests that the neutralization at a layer inside taste cells, taste nerve endings, or the interstices surrounding them is critical in taste transduction.