Differential effects of pertussis toxin on insulin-stimulated phosphatidylcholine hydrolysis and glycerolipid synthesis de novo. Studies in BC3H-1 myocytes and rat adipocytes.
Insulin-induced increases in diacylglycerol (DAG) have been suggested to result from stimulation of de novo phosphatidic acid (PA) synthesis and phosphatidylcholine (PC) hydrolysis. Presently, we found that insulin decreased PC levels of BC3H-1 myocytes and rat adipocytes by approximately 10-25% within 30 s. These decreases were rapidly reversed in both cell types, apparently because of increased PC synthesis de novo. In BC3H-1 myocytes, pertussis toxin inhibited PC resynthesis and insulin effects on the pathway of de novo PA-DAG-PC synthesis, as evidenced by changes in [3H]glycerol incorporation, but did not inhibit insulin-stimulated PC hydrolysis. Pertussis toxin also blocked the later, but not the initial, increase in DAG production in the myocytes. Phorbol esters activated PC hydrolysis in both myocytes and adipocytes, but insulin-induced stimulation of PC hydrolysis was not dependent upon activation of PKC, since this hydrolysis was not inhibited by 500 microM sangivamycin, an effective PKC inhibitor. Our results indicate that insulin increases DAG by pertussis toxin sensitive (PA synthesis de novo) and insensitive (PC hydrolysis) mechanisms, which are mechanistically separate, but functionally interdependent and integrated. PC hydrolysis may contribute importantly to initial increases in DAG, but later sustained increases are apparently largely dependent on insulin-induced stimulation of the pathway of de novo phospholipid synthesis.
Hoffman
J M
JM
James A. Haley Veterans' Hospital, Tampa, Florida 33612.
Standaert
M L
ML
Nair
G P
GP
Farese
R V
RV
D000273:Adipose Tissue / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002460:Cell Line; D002478:Cells, Cultured; D006868:Hydrolysis; D007328:Insulin / Q000494:pharmacology*; D007700:Kinetics; D008297:Male; D009132:Muscles / Q000187:drug effects / Q000378:metabolism*; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism; D037342:Pertussis Toxin; D010712:Phosphatidic Acids / Q000096:biosynthesis*; D010713:Phosphatidylcholines / Q000378:metabolism*; D011741:Pyrimidine Nucleosides / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D007328:Insulin; D009227:Myristic Acids; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D011741:Pyrimidine Nucleosides; D010566:Virulence Factors, Bordetella; D019814:Myristic Acid; C005699:sangivamycin; D037342:Pertussis Toxin; D013755:Tetradecanoylphorbol Acetate