Cell-attached and excised inside-out membrane patches were used to study single channel currents in a cell line derived from human non-pigmented ciliary epithelium. Most of the patches contained a Ca(2+)-dependent K+ channel with large unitary conductance (200 pS in symmetrical K+ solutions). Single channel current in cell-attached patches exposed to high K+ solution in the pipette showed a null potential of -36 mV. This value, which should yield an approximate estimation of cell membrane potential, was reversibly increased by -30 to -40 mV in the presence of Ca2+ ionophores. Tetraethylammonium up to 10 mM applied at the membrane cytoplasmic face had no effect on the channel. Addition of 1 mM BaCl2 to excised patches caused a voltage-dependent blockade of the channel. In the presence of barium the unit currents were not altered, but the channel remained closed for long periods of time and the open state probability decreased with depolarization. The possibility that this channel participates in regulation of transepithelial ciliary body secretion is discussed.