Myosin light chain phosphorylation: modulation of basal and agonist-stimulated venular permeability.
We have previously demonstrated that agonists increase microvascular permeability through a phospholipase C-nitric oxide synthase-guanylate cyclase cascade. The aim of this study was to further investigate the downstream end of the signaling pathway with a focus on myosin light chain (MLC) phosphorylation. The apparent permeability coefficient to albumin was measured in isolated coronary venules. Under control conditions, the nitric oxide donor sodium nitroprusside, as well as the guanosine 3',5'-cyclic monophosphate-dependent protein kinase (PKG) activator 8-bromoguanosine 3',5'-cyclic monophosphate, increased venular permeability two- to threefold. Similarly, activation of protein kinase C (PKC) with phorbol 12-myristate 13-acetate significantly elevated permeability. Inhibition of MLC phosphorylation with ML-7 significantly attenuated the hyperpermeability responses to the agonists. Furthermore, ML-7 dose dependently reduced basal venular permeability. Consistently, inhibition of dephosphorylation with the protein phosphatase inhibitor calyculin dramatically increased basal permeability. These results suggest that 1) PKG and PKC play an important signaling role in the regulation of endothelial barrier function and 2) MLC phosphorylation contributes to basal and agonist-stimulated microvascular permeability.
Yuan
Y
Y
Department of Surgery, Texas A & M University Health Science Center, Temple 76504, USA.
Huang
Q
Q
Wu
H M
HM
D000818:Animals; D001381:Azepines / Q000494:pharmacology; D003326:Coronary Circulation; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000494:pharmacology; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000502:physiology; D018994:Myosin Light Chains / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009281:Naphthalenes / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D010539:Permeability; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013552:Swine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014699:Venules / Q000502:physiology*
D001381:Azepines; D004791:Enzyme Inhibitors; D018994:Myosin Light Chains; D009281:Naphthalenes; C070571:ML 7; D009599:Nitroprusside; C016276:8-bromocyclic GMP; D017869:Cyclic GMP-Dependent Protein Kinases; D011493:Protein Kinase C; D009219:Myosin-Light-Chain Kinase; D010749:Phosphoprotein Phosphatases; D006152:Cyclic GMP; D013755:Tetradecanoylphorbol Acetate