Cyclic GMP modulates the expression of Gi protein and adenylyl cyclase signaling in vascular smooth muscle cells.
We have recently shown that the nitric oxide (NO) donor, SNAP, decreased the expression of Gialpha proteins and associated functions in vascular smooth muscle cells. Because NO stimulates soluble guanylyl cyclase and increases the levels of guanosine 3\',5\'-cyclic monophosphate (cGMP), the present studies were undertaken to investigate whether cGMP can also modulate the expression of Gi proteins and associated adenylyl cyclase signaling. A10 vascular smooth muscle cells (VSMCs) and primary cultured cells from aorta of Sprague Dawley rats were used for these studies. The cells were treated with 8-bromoguanosine 3\',5\'-cyclic monophosphate (8BrcGMP) for 24 h and the expression of Gialpha proteins was determined by immunobloting techniques. Adenylyl cyclase activity was determined by measuring [32P]cAMP formation for [alpha-32P]ATP. Treatment of cells with 8-BrcGMP (0.5 mM) decreased the expression of Gialpha-2 and Gialpha-3 by about 30-45%, which was restored towards control levels by KT5823, an inhibitor of protein kinase G. On the other hand, the levels of Gsalpha protein were not altered by this treatment. The decreased expression of Gialpha proteins by 8Br-cGMP treatment was reflected in decreased Gi functions. For example, the inhibition of forskolin (FSK)-stimulated adenylyl cyclase activity by low concentrations of GTPgammaS (receptor-independent Gi functions) was significantly decreased by 8Br-cGMP treatment. In addition, exposure of the cells to 8Br-cGMP also resulted in the attenuation of angiotensin (Ang) II- and C-ANP4-23 (a ring-deleted analog of atrial natriuretic peptide [ANP])-mediated inhibition of adenylyl cyclase activity (receptor-dependant functions of Gi). On the other hand, Gsalpha-mediated stimulations of adenylyl cyclase by GTPgammaS, isoproterenol and FSK were significantly augmented in 8Br-cGMP-treated cells. These results indicate that 8Br-cGMP decreased the expression of Gialpha proteins and associated functions in VSMCs. From these studies, it can be suggested that 8Br-cGMP-induced decreased levels of Gi proteins and resultant increased levels of cAMP may be an additional mechanism through which cGMP regulates vascular tone and thereby blood pressure.
Bassil
Marcel
M
Department of Physiology, Faculty of Medicine, University of Montreal, Canada.
Anand-Srivastava
Madhu B
MB
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001011:Aorta / Q000378:metabolism; D001794:Blood Pressure; D002227:Carbazoles / Q000494:pharmacology; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism*; D005786:Gene Expression Regulation; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D007211:Indoles / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D051381:Rats; D015398:Signal Transduction
D002227:Carbazoles; D007211:Indoles; C073601:KT 5823; D015124:8-Bromo Cyclic Adenosine Monophosphate; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D000262:Adenylyl Cyclases; D006152:Cyclic GMP