Protein kinases modulate store-operated channels in pulmonary artery smooth muscle cells

• Published in: Journal of biomedical science Vol. 18; no. 1; p. 2
• Main Authors: Chen, I-Shan, Dai, Zen-Kong, Welsh, Donald G, Chen, Ing-Jun, Wu, Bin-Nan
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 06.01.2011; BioMed Central; BMC
• Subjects: Accountants; Adenosine triphosphatase; Animals; Calcium - metabolism; Calcium Channel Blockers - pharmacology; Calcium Channels - metabolism; Cells; Cells, Cultured; Colleges & universities; Female; Inositol; Kinases; Muscular system; Myocytes, Smooth Muscle - cytology; Myocytes, Smooth Muscle - metabolism; Protein Kinase Inhibitors - pharmacology; Protein kinases; Protein Kinases - metabolism; Pulmonary arteries; Pulmonary Artery - cytology; Pulmonary Artery - metabolism; Rats; Rats, Sprague-Dawley; Rodents; Veins & arteries
• ISSN: 1423-0127; 1021-7770; 1423-0127
• DOI: 10.1186/1423-0127-18-2

This study investigates whether protein kinase G (PKG), protein kinase A (PKA) and protein kinase C (PKC) are involved in the regulatory mechanisms of store-operated channel (SOC) in pulmonary arteries. Pulmonary artery smooth muscle cells (PASMCs) were enzymatically dissociated from rat intralobar pulmonary arteries. Whole cell, cell-attached and inside-out patch-clamp electrophysiology were used to monitor SOCs in isolated PASMCs. Initially the Ca2+-ATPase inhibitor cyclopiazonic acid (CPA, 10 μM) initiated a whole cell current that was reduced by the SOC blocker SKF-96365 (10 μM). Subsequent work using both cell-attached and whole cell configurations revealed that the PKG and PKA inhibitors, KT5823 (3 μM) and H-89 (10 μM), also stimulated SOC activity; this augmentation was attenuated by the SOC blockers SKF-96365 (10 μM) and Ni2+ (0.1 mM). Finally using the inside-out configuration, the PKC activator phorbol 12-myristate 13-acetate (PMA, 10 μM) was confirmed to modestly stimulate SOC activity although this augmentation appeared to be more substantial following the application of 10 μM inositol 1,4,5-triphosphate (Ins(1,4,5)P3). SOC activity in PASMCs was stimulated by the inhibition of PKG and PKA and the activation of PKC. Our findings suggest that the SOC could be a substrate of these protein kinases, which therefore would regulate the intracellular concentration of calcium and pulmonary arteriopathy via SOC.