Voltage-dependent inactivation of T-tubular skeletal calcium channels in planar lipid bilayers.
Single-channel properties of dihydropyridine (DHP)-sensitive calcium channels isolated from transverse tubular (T-tube) membrane of skeletal muscle were explored. Single-channel activity was recorded in planar lipid bilayers after fusion of highly purified rabbit T-tube microsomes. Two populations of DHP-sensitive calcium channels were identified. One type of channel (noninactivating) was active (2 microM +/- Bay K 8644) at steady-state membrane potentials and has been studied in other laboratories. The second type of channel (inactivating) was transiently activated during voltage pulses and had a very low open probability (Po) at steady-state membrane potentials. Inactivating channel activity was observed in 47.3% of the experiments (n = 84 bilayers). The nonstationary kinetics of this channel was determined using a standard voltage pulse (HP = -50 mV, pulse to 0 mV). The time constant (tau) of channel activation was 23 ms. During the mV). The time constant (tau) of channel activation was 23 ms. During the pulse, channel activity decayed (inactivated) with a tau of 3.7 s. Noninactivating single-channel activity was well described by a model with two open and two closed states. Inactivating channel activity was described by the same model with the addition of an inactivated state as proposed for cardiac muscle. The single-channel properties were compared with the kinetics of DHP-sensitive inward calcium currents (ICa) measured at the cellular level. Our results support the hypothesis that voltage-dependent inactivation of single DHP-sensitive channels contributes to the decay of ICa.
Mejía-Alvarez
R
R
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas 77030.
Fill
M
M
Stefani
E
E
D000818:Animals; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism*; D004095:Dihydropyridines / Q000494:pharmacology; D066298:In Vitro Techniques; D007700:Kinetics; D008051:Lipid Bilayers; D008564:Membrane Potentials; D008861:Microsomes / Q000378:metabolism; D008954:Models, Biological; D009132:Muscles / Q000378:metabolism*; D011817:Rabbits; D051381:Rats
D015220:Calcium Channels; D004095:Dihydropyridines; D008051:Lipid Bilayers; C038806:1,4-dihydropyridine