Single-electrode voltage-clamp techniques were used to examine membrane currents recorded as a result of hyperpolarizing pre-pulses in cat bladder parasympathetic neurones. In 84 ganglion cells examined, two types of current were observed in response to hyperpolarizing pre-pulses of 10 ms to 1 s duration from holding potentials of -30 to -60 mV to test potentials of -90 to -130 mV. In 46 cells, a short-duration pulse induced a slow inward current (SIC); with longer pulse durations, an outward current was superimposed on the SIC, resulting in a late slow outward current (LSOC). In the remaining cells, either a SIC (n = 12) or an LSOC (n = 26) was recorded over a range of hyperpolarizing pre-pulse durations. The more depolarized the holding potential, the more hyperpolarized the test potential and the longer the pulse duration, the larger the amplitude of the SIC and LSOC. The SIC and LSOC were associated with an increase in input conductance. The extrapolated reversal potential (Vrev) for the LSOC obtained at a holding potential of -60 mV (where the LSOC seemed to be less contaminated with the SIC) was -89 +/- 4 mV (mean +/- standard error of the mean; n = 5), which is close to the equilibrium potential for the K ion. The LSOC was depressed by a high-K (10-20 mM) solution and potentiated by a low-K (0.47 mM) solution. The SIC was depressed by a low-Na (26.2 mM) solution, but was not affected significantly by a low-Cl (12.2 mM) solution. A low-Ca (0.1 mM)/high-Mg (5 mM) solution depressed the LSOC, while a high-Ca (5 mM) solution potentiated it. Cd (0.5 mM) blocked the SIC almost completely, and suppressed the LSOC. The LSOC but not the SIC was suppressed by tetraethylammonium chloride (10 mM). Superfusing Cs (3 mM) did not affect either the LSOC or the SIC. 4-Aminopyridine (1 mM) and muscarine (10 microM) depressed or replaced the SIC with an outward current, while potentiating the LSOC. These results suggest that a hyperpolarizing pre-pulse induces slow inward Na- and late slow outward Ca-dependent K currents, which are inactivated at depolarized potentials and are de-inactivated by hyperpolarizing pulses in a time-dependent manner.