Phosphatidic acid may stimulate membrane receptors mediating adenylate cyclase inhibition and phospholipid breakdown in 3T3 fibroblasts.
Incubation of 3T3 fibroblasts with phosphatidic acid (PA) from egg lecithin or with thrombin resulted in decreases in cellular cAMP due to inhibition of adenylate cyclase, in rapid increases in inositol 1,4,5-tris-,1,4-bis-, and 1-monophosphates probably due to activation of phospholipase C, and in arachidonic acid release. Synthetic PAs consisting of unsaturated fatty acid diesters were as effective as PA from egg lecithin, whereas PAs with saturated fatty acids were only slightly effective and antagonized the effect of active PAs selectively, despite the fact that both types of PA analogues (sodium salts) were apparently dissolved in the incubation medium. PA-induced decreases in cAMP were not affected by omission of Ca2+ from incubation medium but were abolished by prior exposure of cells to islet-activating protein (pertussis toxin). This islet-activating protein treatment of cells was without effect on PA- or thrombin-induced generation of inositol phosphates. Thus, PA-induced inhibition of adenylate cyclase was (but activation of phospholipase C was not) mediated by an islet-activating protein substrate GTP-binding protein. Homologous desensitization was observed with thrombin-, bradykinin-, and PA-induced decreases in cAMP in 3T3 cells; prior exposure of the cells to any one of these agents abolished or greatly diminished the subsequent response to the same agent but did not affect the responses to others. The effects of PA were cell-specific; it failed to decrease cAMP in rabbit platelets in which labeled PA rapidly increasing in response to thrombin or A23187 was mostly outside the cells. Based on these results, it is proposed that PA interacts with its own specific membrane receptors, thereby triggering multiple effector systems in 3T3 cells.
D000067956:Adenylyl Cyclase Inhibitors; D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D001792:Blood Platelets / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D007295:Inositol Phosphates / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D010712:Phosphatidic Acids / Q000494:pharmacology*; D010713:Phosphatidylcholines; D010743:Phospholipids / Q000378:metabolism*; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology
D000067956:Adenylyl Cyclase Inhibitors; D001095:Arachidonic Acids; D002135:Calcium-Binding Proteins; D007295:Inositol Phosphates; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D010743:Phospholipids; D011956:Receptors, Cell Surface; D016718:Arachidonic Acid; D000001:Calcimycin; D000242:Cyclic AMP; D013917:Thrombin