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The Journal of General Physiology, Vol 89, 421-442, Copyright © 1987 by The Rockefeller University Press
ARTICLES |
C Caputo and P Bolanos
Short muscle fibers (approximately 1.5 mm) of Rana pipiens were voltage- clamped with a two-microelectrode technique at a holding potential of - 100 mV. Using conditioning depolarizing ramps, with slopes greater than 0.2 mV/s, partially inactivated responses are obtained at threshold values between -55 and -35 mV. With slopes equal to or slower than 0.1 mV/s, one inactivates contraction without ever activating it. When the membrane potential is brought slowly to values more positive than about -40 mV, test pulses, applied on top of the ramps, bringing the membrane potential to values up to +100 mV, are ineffective in eliciting contractile responses, which indicates complete inactivation. After inactivation, contractile threshold is shifted by perhaps 10 mV, to about -40 mV. The sensitivity of fibers to depolarizing ramps is increased by D-600 (50 microM), dantrolene (50 microM), tetracaine (100 microM), and low calcium (10(-8) M). In the presence of these agents, complete inactivation was obtained using ramp slopes of 1, 0.8, 0.4, and 0.2 mV/s, respectively. Nifedipine was less effective. With D-600, once inactivation had been induced, no repriming occurred after repolarization to -100 mV, and partial recovery occurred after washing out the drug. With low calcium, tetracaine, and nifedipine, the tension- voltage relationship was not affected, whereas the steady state inactivation curve (obtained in repriming experiments) was shifted by 10-25 mV toward more negative potentials. With D-600, the activation curve was not modified, whereas the inactivation curve could not be obtained, because of repriming failure. With dantrolene, the inactivation curve was not affected, whereas the activation curve was shifted toward less negative potentials and peak tension diminished, depending on the pulse duration. The results indicate that it is possible to induce complete inactivation without activation, and to differentiate activation and inactivation parameters pharmacologically, which suggests that the two are separate processes.
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