Abstract
Drosophila mutants provide an ideal model to study channel-type specificity of ion channel regulation in situ. In this study, the effects of divalent cations on voltage-gated K+ currents were investigated in acutely dissociated central neurons of Drosophila third instar larvae using the whole-cell patch-clamp recording. Our data showed that micromolar Cd2+ enhanced the peak inactivating current (IA) without affecting the delayed component (IK). The same results were obtained in Ca2+-free external solution, and from slo1 mutation, which eliminates transient Ca2+-activated K+ current. Micromolar Cd2+and Zn2+, and millimolar Ca2+and Mg2+ all shifted the steady-state inactivation curve of IA without affecting the voltage-dependence of IA activation, whereas millimolar Cd2+markedly affected both the activation and steady-state inactivation curves for IA. Divalent cations affected IA with different potency; the sequence was: Zn2+ > Cd2+ > Ca2+ > Mg2+. The modulation of IA by Cd2+ was partially inhibited in ShM, a null Shaker (one of IA-encoding genes) mutation. Taken together, the channel-type specificity, the asymmetric effects on IA activation and inactivation kinetics, and the diverse potency of divalent cations all strongly support the idea that physiological divalent cations modulate A-type K+ channels through specific binding to extracellular sites of the channels.
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