Calcium currents in isolated rabbit coronary arterial smooth muscle myocytes

Abstract

1. Calcium inward currents were recorded from relaxed enzymatically isolated smooth muscle cells from the rabbit epicardial left descending coronary artery using a single-pipette voltage-clamp technique. Outward K+ currents were blocked with CsCl-tetraethylammonium-filled pipette solutions. 2. Relaxed coronary smooth muscle cells had a maximum diameter of 8.6 +/- 0.6 microns and a cell length of 96.7 +/- 3.3 microns when bathed in 2.5 mM [Ca2+]o. The average resting membrane potential at room temperature was -32 +/- 10 mV. The mean cell capacitance was 18.5 +/- 1.7 pF and the input resistance was 3.79 +/- 0.58 G omega. 3. Depolarizing voltage-clamp steps from a holding potential of -80 mV elicited a single time- and voltage-dependent inward current which was dependent upon extracellular [Ca2+]. In 2.5 mM [Ca2+]o, the inward current was activated at a potential of -40 mV and peaked at +10 mV. This current was inhibited by 0.5 mM-CdCl2 and 1 microM-nifedipine and was enhanced with 1 microM-Bay K 8644. No detectable low-threshold, rapidly inactivating T-type calcium current was observed. 4. The apparent reversal potential of this inward current in 2.5 mM [Ca2+]o was +70 mV and shifted by 33.0 mV per tenfold increase in [Ca2+]o. This channel was also more permeable to barium and strontium ions than to calcium ions. 5. Single calcium channel recordings with 110 mM-Ba2+ as the charge carrier revealed a mean slope conductance of 20.7 +/- 0.8 pS. 6. This calcium current (ICa) exhibited a strong voltage-dependent inactivation process. However, the steady-state inactivation curve (f infinity) displayed a slight nonmonotonic, U-shaped dependence upon membrane potential. The potential at which half of the channels were inactivated was -27.9 mV with a slope factor of 6.9 mV. The steady-state activation curve (d infinity) was also well-described by a Boltzmann distribution with a half-activation potential at -4.4 mV and a slope factor of -63 mV. ICa was fully activated at approximately +20 mV. 7. The rate of inactivation was dependent upon the species of ion carrying the current. Both Sr2+ and Ba2+ decreased the rate as well as the degree of inactivation. The tau f (fitted time constant of inactivation) curve displayed a U-shaped relationship in 2.5 mM [Ca2+]o. The reactivation process was voltage dependent and could be described by a single exponential. 8. The current amplitude and the inactivation kinetics were temperature dependent.(ABSTRACT TRUNCATED AT 400 WORDS)