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Myogenic vasoconstriction in mouse renal interlobar arteries: role of endogenous and ENaC

Department of Physiology and Biophysics and the Center for Excellence in Cardiovascular Renal Research, University of Mississippi Medical Center, Jackson, Mississippi

Submitted 23 May 2006 ; accepted in final form 12 July 2006

Mechanosensitive ion channels are thought to initiate pressure-induced vasoconstriction, however, the molecular identity of these channels is unknown. Recent work from our laboratory suggests that members of the Degenerin/Epithelial Na⁺ Channel (DEG/ENaC) family may be components of the mechanosensitive ion channel complex in vascular smooth muscle (VSM); however, the specific DEG/ENaC proteins mediating myogenic constriction are unknown. The goal of this study is to determine if specific knockdown of or ENaC, using dominant-negative (DN) or small-interference RNA (siRNA) molecules, inhibits pressure-induced vasoconstriction in mouse renal interlobar arteries. To address this goal, isolated arteries were transiently transfected with or ENaC DN-cDNA or siRNA molecules. After 24 h, vessels were either 1) cannulated and pressurized for pressure-diameter response curves or 2) dissociated and immunolabeled to determine VSM cell endogenous ENaC protein expression. We found that transfection of ENaC DN-cDNA or siRNA suppresses -, but not ENaC protein expression. Similarly, ENaC DN-cDNA or siRNA suppresses -, but not ENaC protein expression. In addition, transfection of - or ENaC DN-cDNA or siRNA molecules inhibits pressure-induced vasoconstriction, but does not block agonist-induced vasoconstriction. Our results provide the first direct evidence that and ENaC proteins are essential in mediating myogenic vasoconstriction in mouse renal interlobar arteries.

mechanotransduction; transfected isolated renal vessel; stretch-activated cation channel; siRNA; dominant negative

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