SUMMARY1. Ca2" inward current was studied using the whole-cell patch clamp technique in single smooth muscle cells enzymatically isolated from the rabbit ear artery.Currents were studied in salt solutions containing either normal (1-5 mM) Ca2`or high (110 mM) Ba2+. Outward currents were minimized by using a high-Na+ intracellular solution containing 10 mM-TEA.2. In normal-Ca2+ solution, the threshold at which inward current could be evoked was -60 mV at a holding potential of -80 mV and -48 mV at a holding potential of -60 mV. At both holding potentials, the current showed little inactivation over 500 ms near threshold, and inactivated substantially but incompletely with larger depolarizations. In high-Ba2+ solution where currents were 5-10 times larger, current threshold, maximal peak amplitude and apparent reversal potential were shifted in a positive direction along the voltage axis.3. Inward current in normal-Ca2+ solution was only fully available negative to -90 mV, was half-inactivated near -47 mV and was about 90% inactivated at -10 mV. A component of non-inactivating current was, however, present in both normal-Ca2' and high-Ba2+ solutions even after conditioning pulses of 5 s duration to + 30 mV. The inactivation-potential relationship was shifted in a positive direction in high-Ba2+ solution, and its position showed considerable variation between cells.4. The inward current from a holding potential of -70 mV was reduced in normalCa2+ solution by about 50 % by nifedipine (041 /tM) in some cells at all test potentials, although at more-negative test potentials peak current was unaffected in some cells even at 10 /iM. In some cells when the holding potential was -90 mV, the peak current amplitude was increased markedly by 10 /iM-nifedipine. In high-Ba2+ solution, from negative holding potentials (-70 to -80 mV), nifedipine augmented peak current at test potentials negative to -10 mV and shifted the current threshold in a negative direction; current at potentials positive of -10 mV was reduced. Both effects were concentration dependent. The stimulatory effect of nifedipine was abolished and its inhibitory effect was enhanced when less-negative holding potentials were employed. Bay K 8644 (0-1 or 1 /bM) approximately doubled current * Present address:

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Cellular influx of 24Na was measured in isolated rabbit aorta during stimulation with 10 microM norepinephrine (NE) or depolarization with 80 mM K solution, using a pulse-labeling, cold-wash technique. NE caused a two- to threefold increase in Na influx; a smaller but significant increase was also observed in depolarized tissues. Basal and NE-induced fluxes at 1 min were significantly increased by a 20-min preincubation in a Ca-free solution containing 2 mM EGTA; elevation of [Mg] in this solution reduced these effects. The high K-induced influx was prevented by a combination of low Ca (30 microns) and elevated Mg (10 mM). The Ca agonist, BAY-K 8644, increased 24Na influx. The Ca antagonist, diltiazem, inhibited the depolarization-stimulated 24Na influx in a concentration-dependent manner, but was less effective in blocking the response to NE. Extension of the preincubation in NE plus Ca-free medium from 30 s to 15 min decreased the influx response and contraction. After exposure to NE in Ca-free solution, 24Na influx remained elevated 10 min after washing out NE in the continued absence of Ca. A second exposure to NE at that time did not increase influx. We propose that a component of 24Na influx during excitation depends directly on a rise in intracellular [Ca]. The role of an indirect effect of [Ca] on metabolic H+ production with subsequent stimulation of the Na+-H+ exchange may also be a factor.

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