Molecular mechanisms of homologous and heterologous desensitization mediated by vasopressin in smooth muscle cells.
Arginine vasopressin (AVP) has been shown previously to enhance phosphatidylinositol (PI) turnover and mobilize calcium in the rat aortic smooth muscle cell-line (A10; ATCC CRL 1476) via the V1 receptor (Aiyar, N., Nambi, P., Stassen, F. L., and Crooke, S. T. (1986) Life Sci. 39, 37-45). Exposure of A10 cells to AVP for periods ranging from 5 min to 2 h resulted in 30-40% loss in AVP-binding sites and an inhibition of the production of inositol di- and trisphosphates and the mobilization of calcium when the cells were rechallenged by addition of AVP. We now report that during the same time course AVP induces a dose- and time-dependent decrease in labeled PI, phosphatidylinositol-4-phosphate and phosphatidylinositol-4,5-bisphosphate reaching a minimum after 30 min of incubation. After 2 h of exposure to AVP, the levels of labeled PI, phosphatidylinositol-4-phosphate and phosphatidylinositol-4,5-bisphosphate increased to a new basal level approximately 30% less than the untreated cultures. The decrease in inositol lipid labeling mediated by AVP was inhibited when the V1 antagonist SK&F 100273 was included in the incubations with AVP. No decrease was observed when the V2 agonist 1-deamino, [8-D-arginine]vasopressin was used for pretreatment of the cells. Furthermore, when PI kinase activity was measured in cell extracts from untreated and AVP-treated (2 h) cells a significant decrease (p less than 0.05) was observed in the absence, but not in the presence, of added PI in the AVP-treated cells as compared with the control cells. Thrombin also stimulates PI metabolism and calcium mobilization in these cells and brought about both a prolonged decrease in inositol lipids and inhibition of PI kinase activity. AVP pretreatment affected the release of intracellular Ca2+ induced by AVP, thrombin, and ATP, differently. The time of AVP pretreatment required to induce half-maximal inhibition of intracellular Ca2+ release in response to AVP, thrombin, and ATP was approximately 8, 24, and 30 min, respectively. Consequently, we suggest that the reduction in response to AVP with short term preincubation is due to homologous desensitization as reflected by 30-40% decrease in V1 receptors. Subsequently, a decrease in inositol lipid pools and PI kinase activity results in heterologous desensitization in response to AVP, thrombin, and ATP.
Grier
C E
CE
Smith Kline and French Laboratories, Department of Molecular Pharmacology, King of Prussia, Pennsylvania 19406-0939.
Nambi
P
P
Aiyar
N
N
Crooke
S T
ST
D019870:1-Phosphatidylinositol 4-Kinase; D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism; D001127:Arginine Vasopressin / Q000494:pharmacology*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003894:Deamino Arginine Vasopressin / Q000494:pharmacology*; D007700:Kinetics; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000378:metabolism; D010770:Phosphotransferases / Q000378:metabolism; D051381:Rats; D013917:Thrombin / Q000502:physiology
D010716:Phosphatidylinositols; D001127:Arginine Vasopressin; D010770:Phosphotransferases; D019870:1-Phosphatidylinositol 4-Kinase; D013917:Thrombin; D003894:Deamino Arginine Vasopressin