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TTX-sensitive voltage-gated Na⁺ channels are expressed in mesenteric artery smooth muscle cells

Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland

Submitted 17 November 2004 ; accepted in final form 11 February 2005

The presence and properties of voltage-gated Na⁺ channels in mesenteric artery smooth muscle cells (SMCs) were studied using whole cell patch-clamp recording. SMCs from mouse and rat mesenteric arteries were enzymatically dissociated using two dissociation protocols with different enzyme combinations. Na⁺ and Ca²⁺ channel currents were present in myocytes isolated with collagenase and elastase. In contrast, Na⁺ currents were not detected, but Ca²⁺ currents were present in cells isolated with papain and collagenase. Ca²⁺ currents were blocked by nifedipine. The Na⁺ current was insensitive to nifedipine, sensitive to changes in the extracellular Na⁺ concentration, and blocked by tetrodotoxin with an IC₅₀ at 4.3 nM. The Na⁺ conductance was half maximally activated at –16 mV, and steady-state inactivation was half-maximal at –53 mV. These values are similar to those reported in various SMC types. In the presence of 1 µM batrachotoxin, the Na⁺ conductance-voltage relationship was shifted by 27 mV in the hyperpolarizing direction, inactivation was almost completely eliminated, and the deactivation rate was decreased. The present study indicates that TTX-sensitive, voltage-gated Na⁺ channels are present in SMCs from the rat and mouse mesenteric artery. The presence of these channels in freshly isolated SMC depends critically on the enzymatic dissociation conditions. This could resolve controversy about the presence of Na⁺ channels in arterial smooth muscle.

vascular smooth muscle; freshly isolated cells; whole cell voltage clamp; batrachotoxin

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