Inactivation of L-type Ca channels in embryonic chick ventricle cells: dependence on the cytoskeletal agents colchicine and taxol

• Published in: Biophysical journal Vol. 67; no. 6; pp. 2296 - 2304
• Main Authors: Galli, A., DeFelice, L.J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-12-1994; The Biophysical Society
• Subjects: Animals; Biophysical Phenomena; Biophysics; Calcium Channels - classification; Calcium Channels - drug effects; Calcium Channels - metabolism; Chick Embryo; Colchicine - pharmacology; Cytoskeleton - drug effects; Cytoskeleton - metabolism; Heart Ventricles - drug effects; Heart Ventricles - embryology; Heart Ventricles - metabolism; In Vitro Techniques; Kinetics; Models, Biological; Paclitaxel - pharmacology
• ISSN: 0006-3495; 1542-0086
• DOI: 10.1016/S0006-3495(94)80715-5

This article shows that colchicine and taxol strongly influence the kinetics of L-type Ca channels in intact cardiac cells, and it suggests a mechanism for this action. It is known that colchicine disassociates microtubules into tubulin, and that taxol stabilizes microtubules. We have found that colchicine increases the probability that Ca channels are in the closed state and that taxol increases the probability they are in the open state. Moreover, taxol lengthens the mean open time of Ca channels. In this regard, taxol is similar to Bay-K 8644; however, Bay K works on inside-out patches, but taxol does not. Neither colchicine nor taxol alters the number of Ca channels in a patch. We have quantified these results as follows. It is known that L-type channels in embryonic chick heart ventricle cells have voltage- and current-dependent inactivation. In 10 mM Ba, channel conductance is linear in the range -10 to 20 mV. The conductance is 12 +/- 1 pS, and the extrapolated reversal potential is 42 +/- 2 mV (n = 3). In cell-attached patches, inactivation depends on the number of channels. One channel (holding at -80 mV and stepping to 0 mV for 500 ms) shows virtually no inactivation. However, three channels inactivate with a time constant of 360 +/- 20 ms (n = 6). In similar patches, colchicine (80 microM for 15 min) decreases the inactivation time constant to 162 +/- 33 ms (n = 4) and taxol (50 microM for 10 min) virtually abolishes inactivation (time constant 812 +/- 265 ms (n = 4)).