The regulation of voltage-activated K(+), and Ca(2+) currents by actin filaments was studied in salamander retinal ganglion cells, using the whole-cell patch clamp technique and Ca(2+) imaging. Disruption of F-actin by cytochalasin B or latrunculin B resulted in a reduction of L-type Ca(2+) current by 55+/-4%, and a sustained outward K(+) current (I(k)) by 41+/-3%. The effect was diminished when the F-actin stabilizing agent phalloidin was present in the patch pipette. In a group of cells where I(K) exhibited a small degree of inactivation, the effect of F-actin disruption on current was dual; it increased it by 89+/-16%, at -10 mV, and reduced it by 37+/-5% at +50 mV voltage step from the same holding potential of -70 mV. This was accompanied by a shift in a voltage of half-maximal activation toward negative potentials by approximately 20 mV. In Ca(2+) imaging experiments, 30 min incubation of isolated neurons with latrunculin A reduced a depolarization-induced Ca(2+) accumulation by 45+/-5%. These results suggest a role for the actin cytoskeleton in regulating voltage-gated ion channels in retinal ganglion cells.