© 2000 American Heart Association, Inc.
Cellular Biology |
Microtubule Disruption Modulates Ca2+ Signaling in Rat Cardiac Myocytes
From Physiopathologie Cardiovasculaire, INSERM U-390, Montpellier, France.
Correspondence to Ana M. Gómez, INSERM U-390, CHU Arnaud de Villeneuve, 34295 Montpellier, France. E-mail agomez{at}u390.montp.inserm.fr
Abstract—Microtubules have been shown to alter contraction in cardiac myocytes through changes in cellular stiffness. However, an effect on excitation-contraction coupling has not been examined. Here we analyze the effects of microtubule disruption by 1 µmol/L colchicine on calcium currents (ICa) and [Ca2+]i transients in rat ventricular myocytes. ICa was studied using the whole-cell patch-clamp technique. Colchicine treatment increased ICa density (peak values, -4.6±0.4 and -9.1±1.3 pA/pF in 11 control and 12 colchicine-treated myocytes, respectively; P<0.05). ICa inactivation was well fitted by a biexponential function. The slow component of inactivation was unchanged, whereas the fast component was accelerated after colchicine treatment (at -10 mV, 11.8±1.0 versus 6.7±1.0 ms in control versus colchicine-treated cells; P<0.005). [Ca2+]i transients were analyzed by fluo-3 epifluorescence simultaneously with ICa. Peak [Ca2+]i transients were significantly increased in cardiac myocytes treated with colchicine. The values of F/F0 at 0 mV were 1.1±0.02 in 9 control cells and 1.4±0.1 in 11 colchicine-treated cells (P<0.05). ß-Adrenergic stimulation with 1 µmol/L isoproterenol increased both ICa and [Ca2+]i transient in control cells. However, no significant change was induced by isoproterenol on colchicine-treated cells. Colchicine and isoproterenol effects were similar and not additive. Inhibition of adenylyl cyclase by 200 µmol/L 2'-deoxyadenosine 3'-monophosphate blunted the colchicine effect. We suggest that ß-adrenergic stimulation and microtubule disruption share a common pathway to enhance ICa and [Ca2+]i transient.
Key Words: heart • Ca2+ current • microtubule • Ca2+ transient • ß-adrenergic stimulation
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