title: Extracellular K+ activates a K(+)- and H(+)-permeable conductance in frog taste receptor cells.
abstract: 1. The effect of extracellular K+ on membrane currents of bull frog (Rana catesbeiana) taste receptor cells (TRCs) was investigated by the patch clamp and fast perfusion techniques. Extracellular K+ (2.5-90 mM) increased a TRC resting conductance and enhanced both inward and outward whole-cell currents. 2. To isolate the inward current activated by external potassium (PA current), TRCs were dialysed with 110 mM NMGCl while extracellular NaCl was replaced with NMGCl. Under these conditions, the PA current displayed an S-shaped current-voltage (I-V) curve in the -100 to 100 mV range. Extracellular Rb+ and NH4+, but not Li+, Na+ or Cs+, evoked similar currents. 3. The PA current reversal potential (Vr) did not follow the equilibrium K+ potential under experimental conditions. Therefore, K+ ions were not the only current carriers. The influence of other ions on the PA current Vr indicated that the channels involved are permeable to K+ and H+ and much less so to Na+, Ca2+ and Mg2+. Relative permeabilities were estimated on the basis of the Goldman-Hodgkin-Katz equation as follows: PH:PK:PNa = 4000:1:0.04. 4. All I-V curves of the PA current were nearly linear at low negative potentials. The slope conductance at these voltages was used to characterize the dependence of the PA current on external K+ and H+. The slope conductance versus K+ concentration was fitted by the Hill equation. The data yielded a half-maximal concentration, K1/2 = 19 +/- 3 mM and a Hill coefficient, nH = 1.53 +/- 0.36 (means +/- S.E.M.). 5. The dependence of the mean PA current and the current variance on the K+ concentration indicated a rise in the open probability of the corresponding channels as extracellular K+ was increased. With 110 mM KCl in the bath, the single channel conductance was estimated at about 6 pS. Taken together, the data suggest that extracellular K+ may serve as a ligand to activate specific small-conductance cation channels (PA channels). The mean number of the PA channels per TRC was estimated as at least 2000. 6. Extracellular Ba2+, Cd2+, Co2+, Ni2+ and Cs+ blocked the PA current in a potential-dependent manner. The PA current was blocked by Cs+ as quickly as the blocker could be applied (approximately 15 ms). The time course of the divalent cation block was well fitted by a single exponential function. The time constants were estimated at 26.5 +/- 1.9, 41.7 +/- 3.1, 56.1 +/- 4.2 and 370 +/- 18 ms at 1 mM Cd2+, Co2+, Ni2+ and Ba2+, respectively. The blocker efficiency at negative voltages followed the sequence: Cs+ > Cd2+ > Ba2+ > Ni2+ > Co2+. 7. The data indicate that protons and divalent blockers act within the PA channel pore and that H+ and the divalent ions probably act via similar mechanisms to affect the PA current. These observations and the strong pH dependence of the PA current Vr suggest that H+ occupation of the PA channel pore leading to interruption of K+ flux is the main mechanism of the pH dependence of the PA current. 8. Extracellular K+ enhanced the sensitivity of isolated TRCs to bath solution acidification due to activation of the PA channels. With 10 mM K+ in the bath, half-maximal depolarization of the TRCs was observed at pH values of 6.4-6.8. The possible role of the PA channels in sour transduction is discussed.
authors.affiliation: Howard Hughes Medical Institute, New York, NY, USA.
authors.forename: S S
authors.initials: SS
authors.lastname: Kolesnikov
authors.forename: R F
authors.initials: RF
authors.lastname: Margolskee
mesh_terms: D000818:Animals; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D004594:Electrophysiology; D005110:Extracellular Space / Q000378:metabolism; D006859:Hydrogen / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009431:Neural Conduction / Q000187:drug effects; D018408:Patch-Clamp Techniques; D010477:Perfusion; D011188:Potassium / Q000494:pharmacology*; D015221:Potassium Channels / Q000819:agonists* / Q000187:drug effects; D011892:Rana catesbeiana; D013650:Taste Buds / Q000187:drug effects / Q000378:metabolism*
chemical_list: D015221:Potassium Channels; D006859:Hydrogen; D011188:Potassium