Abstract
Regulation of the Cystic Fibrosis Transmembrane Conductance Regulator Channel by β-Adrenergic Agonists and Vasoactive Intestinal Peptide in Rat Smooth Muscle Cells and Its Role in Vasorelaxation
Highlights
The balance between contraction and relaxation signals determines the vascular tone, the major determinant of the resistance to blood flow through the circulation
These results demonstrated that CFTR endogenously expressed in smooth muscle cells (SMC) can be phosphorylated in vitro by PKA as in epithelial cells [10, 11, 18]
1) Activation of -adrenergic or vasoactive intestinal peptide (VIP) receptors and subsequently of the adenylyl cyclasecAMP-PKA phosphorylation pathway leads to activation of CFTR in SMC. 2) Pharmacological activation/inhibition of CFTR in SMC is similar to that of the epithelial CFTR. 3) CFTR activation in SMC leads to vasorelaxation
Summary
The thoracic aorta of animals killed by cervical dislocation was removed and placed into Krebs solution containing 120 mM NaCl, 4.7 mM KCl, 2.5 mM CaCl2, 1.2 mM MgCl2, 15 mM NaHCO3, 1.2 mM KH2PO4, 11 mM D-glucose, 10 mM Hepes, pH 7.4. SMC Isolation—For each culture, thoracic aortas from four rats were excised and placed in modified Krebs solution containing 120.8 mM NaCl, 5.9 mM KCl, 0.2 mM CaCl2, 1.2 mM MgCl2, 1.2 mM NaH2PO4, 2 mM NaHCO3, 11 mM D-glucose, 10 mM Hepes, and pH 7.4. Cell lysates were supplemented with 3 volumes of NET buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 5 mM EDTA, 0.05% Nonidet P-40) and incubated overnight at 4 °C with 2 g of CFTR mouse monoclonal antibody clone 24 –1 (R & D Systems) or 2 g of nonimmune mouse IgG (Sigma). Data are presented as mean Ϯ S.E. of n experiments
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