The patch-clamp recording technique was used to examine the properties of the K+ channels in cultured equine sweat gland epithelial cells. With symmetric K+ solutions (140 mM), a single population of K+ channels was identified with a slope conductance of 187 pS and a reversal potential of around 0 mV. The channel was selective for K+ over Na+. Channel activity was increased by membrane depolarization. A 10-fold increase in [Ca2+]i produced an approximate 60 mV negative shift in the open state probability (Popen)-voltage curve. Externally applied tetraethylammonium ions (TEA+) caused a rapid and flickery block of the channel and reduced the unitary current amplitude. TEA+ bound to the blocking site with stoichiometry of 1:1 and with a dissociation constant (Kd) of 186 ± 27 µM at +40 mV. A weak voltage dependence of Kd was observed. Iberiotoxin (100 nM) reduced Popen but had no effect on single-channel conductance. Neither glibenclamide (10 µM) nor intracellular adenosine 5'-triphosphate (ATP, 1 mM) altered channel activity. In addition, ATP, when applied extracellularly, transiently activated the channel by increasing Popen. Channel activity was low around the resting membrane potential in the intact epithelia, indicating that these channels might not contribute to the resting K+ conductance. However, the channel could be activated in a regulated manner. The K+ channels may play a role in transepithelial fluid secretion in sweat gland.