Muscarinic activation and calcium permeation of non-selective cation currents in airway myocytes. Fleischmann, Bernd K., Yong-Xiao Wang, Michael I. Kotlikoff. Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104-6046
APStracts 3:0272C, 1996.
We examined the activation and calcium permeation of non-selective cation channels in voltage-clamped (nystatin), fura 2 -loaded equine tracheal myocytes at 35 degrees C. Methacholine (mACh, 50 [mu]M) induced a biphasic increase in [Ca++]i and a biphasic inward current consisting of a large, rapidly inactivating calcium-activated chloride current (I(Cl)Ca) and a smaller, sustained non-selective cation current (Icat). I(Cl)Ca but not ICat was activated by caffeine. Neither ICat nor the sustained rise in [Ca++]i were blocked by nisoldipine, whereas both were rapidly blocked by Ni++. ICat was determined to be calcium permeant since: 1) a sustained elevation of [Ca++]i occurred when ICat was activated, and blockade of ICat produced a rapid decline in [Ca++]i; 2) increasing extracellular Ca++ during ICat increased [Ca++]i; 3) 110 mM extracellular Ca++ shifted the reversal potential of ICat to 12 mV (Ca++/Cs+ permeability ratio = 3.6); 4 instantaneous voltage-clamp steps to negative potentials during ICat increased the current and [Ca++]i, whereas depolarizing steps decreased the current and [Ca++]i. The fraction of ICat carried by calcium under physiological conditions was estimated to be 14% at -60 mV.

Received 6 June 1996; accepted in final form 7 August 1996.
APS Manuscript Number C319-6.
Article publication pending Am. J. Physiol. (Cell Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 29 August 1996