Single smooth muscle cells isolated from the urinary bladder of the guinea-pig were studied at 35 degrees C in a solution composed of 150 mM NaCl, 3.6 mM CaCl2, 1.2 mM MgCl2, 5.4 mM KCl, 20 mM TEA-Cl, 5 mM glucose, 10 mM HEPES/NaOH (pH 7.4). Whole cells were clamped with a single patch electrode. The clamp settled a step from -65 to -5 mV within 260 microseconds, and afterwards the voltage inhomogeneities were less than 2 mV (measured at the cell edge with a second electrode). The calcium inward current iCa was dissected from net currents by blocking potassium outward currents by means of patch electrodes filled with 130 mM CsCl (Klöckner and Isenberg 1985 a). Pyruvate, succinate and oxalacetate in the patch electrode stabilized iCa and prevented its "run down". 140 ms long clamp steps from -65 to -5 mV evoked a net inward current which could be reversibly blocked by 5 mM NiCl2. The "Ni-sensitive" difference current iCa peaked within 2-4 ms to about 1 nA per cell. Afterwards it completely inactivated; the inactivation could be fitted with three exponentials (time constants of 4, 30, and 250 ms, respectively). The half decay time of 16 ms suggests that most of the inactivation resulted from the fast exponential process. The reference current in the presence of Ni was nearly time independent and almost zero; therefore, iCa could be approximated from the net inward current using the zero current as a reference line.(ABSTRACT TRUNCATED AT 250 WORDS)