Characterization of TWIK-2,a Two-Pore Domain K + Channel,Cloned from the Rat Middle Cerebral Artery

Abstract

TWIK-2, a member of the Two-Pore Domain K channel family, is expressed in a number of mammalian tissues including the vascular system. The function of TWIK-2 is not known. The purpose of this study was to clone the TWIK-2 channel from the rat middle cerebral artery, express it in CHO cells, and characterize the channel’s electrical properties. In light of the fact that there are no specific TWIK-2 inhibitors or activators, a better characterization of the channel should enhance our understanding of its role in the vascular system. TWIK-2 was cloned from the rat middle cerebral artery and expressed with an N-terminal green fluorescence protein (GFP) in CHO cells. We report that rTWIK-2-GFP currents were relatively linear at physiological K⁺ concentrations but become slightly inwardly rectifying in symmetrical K⁺. rTWIK-2-GFP was insensitive to 10 mM TEA, 3 mM 4-aminopyridine, and 10 μM glibenclamide. However, rTWIK-2-GFP was inhibited by Ba²⁺ with 50% of the current being blocked at 80 μM. rTWIK-2-GFP activity was enhanced 60% by 100 μM arachidonic acid. The electrophysiological characteristics of TWIK-2 indicate that it could serve an important role in ion homeostasis and regulation of the membrane potential in arteries and arterioles.

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