Dual effects of dihydropyridines on whole cell and unitary calcium currents in single ventricular cells of guinea-pig

Abstract

We studied the effects of dihydropyridine Ca channel ligands (DHPs), mainly nitrendipine and Bay K8644, on whole cell and single channel Ca currents on single myocytes isolated from the adult guinea-pig ventricle. Nitrendipine had dual effects, stimulatory or inhibitory, depending upon the membrane potential. At low frequencies (less than 0.03 Hz) and negative holding potentials (-90 mV or more), nitrendipine increased the Ca currents in a dose-dependent manner. The dose-response curve was best fitted by a Langmuir adsorption isotherm model which was the sum of two independent one-to-one drug-receptor sites with median effective doses (ED50S) of 1.0 X 10(-9) M and 1.4 X 10(-6) M respectively. When the membrane potential was held at -30 mV or less, nitrendipine inhibited the Ca currents, also in a dose-dependent manner. The dose-response curve was fitted by a single binding site model having a median inhibitor concentration (IC50) of 1.5 X 10(-9) M. At holding potentials between -70 and -40 mV, nitrendipine produced mixed effects on Ca currents; an increase occurred initially and this was followed by a decrease. When rundown was excluded, Bay K8644 showed only stimulatory effects on the Ca currents between holding potentials of -120 and -30 mV. When the test potential was zero or +10 mV the Ca currents reached peak values and the dose-response curve was best fitted by a single binding site model having an ED50 of 3 X 10(-8) M. When the effects were measured at negative test potentials of -30 to -10 mV, the curve was best fitted by a two-site model with ED50S of 3 X 10(-9) and 9 X 10(-7) M. At the single Ca channel level the stimulatory effect of nitrendipine was due to an increased probability that a Ca channel which had opened once would reopen, a reduction in records without activity and an increase in the mean open time. There were no changes in unit conductance. Inhibitory effects were due to a large increase in nulls. At lower concentrations the main effect of Bay K8644 was an increase in the probability of opening. At doses above 10(-6) M, a pronounced increase in the open time was observed. The effects we observed are attributed to at least two sites for DHP related to Ca channels; one with high affinity and one with a lower affinity. The low affinity site mediates a stimulatory effect due to greatly prolonged openings.(ABSTRACT TRUNCATED AT 400 WORDS)