1. Transmembrane pH gradients (acidic inside) and electrical gradients (negative inside) were estimated in cortical synaptosomes from the distribution of the weak base methylamine and the lipophilic cation tetraphenylphosphonium, respectively.2. Acidic interior pH gradients were produced by outwardly directed K + gradients in Na + -free media. External K' accelerated the dissipation of preformed H + gradients. The appearance of H + in the medium was directly demonstrated by pH-stat titration of a weakly buffered medium. Amiloride failed to inhibit K+-induced H + release.3. Elevating K + in the absence of Na' did not affect the endogenous contents of noradrenaline, dopamine, and serotonin, as determined by high-performance liquid chromatography with electrochemical detection.4. H + diffusion potentials were generated when outwardly directed H + gradients were imposed onto the plasma membrane indicating an electrogenic H + efflux which is not coupled to other ions.5. At low K + in the Na+-free sucrose medium, the plasma membrane potential Em (derived from distribution of tetraphenylphosphorium cation) did not approach a value for EK, the K + equilibrium potential (calculated from K + gradients). The deviation of Em from EK could be quantitatively described by a modified constant-field equation, taking a relative H + / K + permeability coefficient of 12400 into consideration.6. It is concluded that synaptosomes have a H + conductance pathway in their plasma membrane in addition to the N a + / H + antiporter. H + influx is driven by and leads to a reduction of Em. K + / H + exchange resulted from the electrical coupling of K + and H + fluxes via parallel K + and H + channels. Since the Na+/H+ antiporter counteracts passive equilibration of H As pointed out by Busa and Nuccitelli [I], aerobic metabolism does not impose an intracellular acid load and hence does not require a pHi-regulating system. However, an acidifying effect results from the passive entry of H + into the cell by diffusion down the H + electrochemical gradient [l]. Because of the low concentrations of H + in biological systems, H + conductance of plasma membranes is generally assumed to be negligibly low. In contrast, a number of recent studies revealed that at least some biological membranes exhibit significant H + permeability, for instance those of sarcoplasmic reticulum [5], kidney brush border vesicles [6 -91, and Ehrlich ascites tumor cells [lo].