P2-purinoceptor-mediated membrane currents in DDT1 MF-2 smooth muscle cells

The electrophysiological response evoked by ATP was investigated in the DDT1 MF-2 smooth muscle cell line using the microelectrode technique and the whole-cell patch clamp technique. Application of ATP (10(-3) M) to the bathing solution caused a small initial depolarization of the cell membrane, followed by hyperpolarization and slow depolarization. During voltage clamping (-50 mV) a triphasic response was recorded on stimulation with ATP (10(-4)-10(-3) M). A short-lasting inward current was followed by a transient outward current and a slowly decreasing inward current. This response was not affected by the receptor antagonists, propranolol (3 X 10(-6) M), phentolamine (3 X 10(-6) M), atropine (3 X 10(-6) M) or theophylline (10(-3) M). The ATP-induced currents were not modified by the voltage-dependent channel blocking agents, tetraethyl ammonium (3 X 10(-3) M), 3,4-diaminopyridine (10(-3) M), tetrodotoxin (3 X 10(-7) M) or diltiazem (10(-5) M). The fast inward current was not detectable at a low ATP concentration (10(-5) M). The outward current showed a reversal potential near -76 mV, which equals the potassium equilibrium potential. This current was abolished after neutralization of the potassium electrochemical gradient. The outward current was suppressed under calcium-free conditions and also in the presence of tolbutamide (10(-4) M) or glipizide (5 X 10(-6) M). Guanosine triphosphate (5 X 10(-6) M) promoted the outward current, while this current was inhibited in the presence of guanosine diphosphate (5 X 10(-6) M).(ABSTRACT TRUNCATED AT 250 WORDS)