A potential-dependent fast outward current in single smooth muscle cells isolated from the newborn rat ileum.

Abstract

1. Whole-cell outward currents have been studied in single smooth muscle cells isolated from newborn and adult rat ileum, using fire-polished glass micropipettes. 2. Two major outward currents, delayed (I(do)) and fast inactivating potential-dependent (I(fo)), have been observed in the newborn rat ileal cells. I(fo) is activated between -50 and -40 mV from the holding potential of -80 mV, whereas I(do) usually starts to activate at membrane potentials positive to -20 mV. Activation of I(do) was fast, its time-to-peak decreased from 10.8 +/- 0.9 ms (n = 5) at -30 mV to 4.5 +/- 0.7 ms (n = 4) at 20 mV. 3. I(fo) decay was monoexponential and its time constant did not depend on the membrane potential. Dependence of I(fo) inactivation on membrane voltage in normal physiological salt solutions (PSS) could be described by the Boltzmann equation with the following parameters: a half-inactivation potential, V0.5 = -70.8 mV and slope factor, k = 7.7 mV. 4. Recovery of I(fo) from inactivation was fitted by a single exponential and was potential dependent. The average time constant was 28.4 +/- 2.4 ms (n = 11) at -120 mV, 47.7 +/- 3.0 ms (n = 6) at -100 mV and 89.6 +/- 5.3 ms (n = 13) at -80 mV. 5. Removal of Ca2+ ions from the PSS (in the presence of 5 mM-Mg2+) increased I(fo) amplitude by about two times, and shifted its voltage dependence of inactivation towards negative membrane potentials by about 16 mV (V0.5 = -87.2 mV). Removal of Mg2+ from the PSS (in the presence of 2.5 mM-Ca2+) exerted no effects upon either inactivation dependence (V0.5 = -74.2 mV) or I(fo) amplitude. 6. I(do) and I(fo) had different sensitivities to K+ channel blockers. With 10 mM-external TEA+ I(do), was preferentially suppressed, while 5 mM-4-aminopyridine (4-AP) completely blocked I(fo). I(fo) was also partially blocked by a higher TEA+ concentration (30 mM), which suppressed I(fo) to 0.55 +/- 0.02 (n = 9). The blocking effect of 4-AP on I(fo) was potential, use and time dependent. 7. Ileal cells isolated from the adult rat demonstrated the presence of two populations of smooth muscle cells. One has an outward current which seems to be similar to that described in the newborn rat. However, in other cells spontaneous transient outward currents, well described in other single smooth muscle cells, but not found in newborn rat ileal cells, have been observed.