Mechanosensitive ion channels have been described in many types of cells. These channels are believed to transduce pressure signals into intracellular biochemical and physiological events. In this study, the patch-clamp technique was used to idendfy and characterize a mechanosensitive ion channel in rat atrial cells. In cell-attached patches, negative pressure in the pipette activated an ion channel in a pressure-dependent manner. The pressure to induce half-maximal activation was 12 -3 mmHg at +40 mV, and nearly full activation was observed at ~20 mmHg. The probability of opening was voltage dependent, with greater channel activity at depolarized potentials. The mechanosensitive channel was identical to the K + channel previously shown to be activated by arachidonic acid and other lipophilic compounds, as judged by the outwardly rectifying current-voltage relation, single channel amplitude, mean open time (1.4 +-. 0.3 ms), bursty openings, K + selectivity, insensitivity to any known organic inhibitors of ion channels, and pH sensitivity. In symmetrical 140 mM KC1, the slope conductance was 94 -+ 11 pS at +60 mV and 64 -8 pS at -60 mV. Anions and cations such as CI-, glutamate, Na +, Cs +, Li +, Ca 2+, and Ba 2+ were not permeant. Extracellular Ba ~+ (1 raM) blocked the inward K + current completely. GdCI3 (100 tiM) or CaCI2 (100 wM) did not alter the K + channel activity or amplitude. Lowering of intracellular pH increased the pressure sensitivity of the channel. The K + channel could be activated in the presence of 5 mM intracellular [ATP] or 10 IzM glybenclamide in inside-out patches. In the absence of ATP, when the ATP-sensitive K + channel was active, the mechanosensitire channel could further be activated by pressure, suggesting that they were two separate channels. The ATP-sensitive K + channel was not mechanosensitive. Pressure activated the K + channel in the presence of albumin, a fatty acid binding protein, suggesting that pressure and arachidonic acid activate the K + channel via separate pathways.