Changes in the surface pH of voltage-clamped snail neurones apparently caused by H+ fluxes through a channel

Abstract

1. The surface and intracellular pH of snail neurones was recorded with microelectrodes while the membrane potential was reduced in 10 mV steps for a few seconds each or to positive values for periods of several minutes. 2. Depolarizations to positive membrane potentials caused rapid falls in surface pH (pHs) which varied from cell to cell and from one point to another on the surface of the same cell. 3. When pHi was normal or alkaline, the first few 10 mV steps of depolarization often caused a small pHs increase which changed to a decrease as the depolarization increased. The threshold potential at which the pHs increase changed to a decrease varied with pHi in a linear manner, so that at acid pHi values the threshold potential approached the normal resting potential. There was good agreement between the threshold and H+ equilibrium potentials calculated from pHi and pHs. 4. The size of the pHs decrease observed at a given pHi and depolarization depended on extracellular buffering power in a non-linear manner. Solutions buffered with 20 mM-NaHCO3 had similar surface buffering power to CO2-free solutions buffered with only 1-2 mM-HEPES, pH 7.5. 5. In 1 mM-HEPES pHs changes were larger, and pHi increases slower, than those seen in cells depolarized to the same potential in 20 mM-HEPES. The slowing of the rate of pHi increase suggests that the pHs changes occur all over the cell surface, and not only at the recording site. 6. With long-lasting depolarizations the size of the pHs decrease was proportional to the rate of pHi increase and thus, assuming a constant intracellular buffering power, to the rate of efflux of H+. 7. The results provide further evidence that snail neurones possess a channel permeable to H+ which is opened on depolarization. H+ efflux through this channel could cause rapid acidification of a confined extracellular space.