Response of 3T3 cells on stimulation of adenylate cyclase by forskolin, studied by multinuclear NMR spectroscopy.
Stimulation of 3T3 cells by forskolin (30 microM) led to an increase in cellular cAMP levels, as detected by 31P-NMR, in both PCA extracts and living cells incorporated in basement membrane gel (BMG) threads. At the same time cAMP was exported in large amounts to the extracellular space. The in vivo experiments showed a decrease of more than 50% in both the PCr and ATP levels within 90 min of stimulation with forskolin. Reperfusion with fresh medium without the drug resulted in the full recovery of the phosphorus energy metabolism. A significant increase in phosphocholine (PC) was observed in 1H and 31P spectra of the PCA extracts concomitant to the rise in cAMP level. 13C spectra of PCA extracts prepared after forskolin stimulation in the presence of [1-13C]glucose indicated a serious inhibition of glycolysis, which was followed by a disordering of the phosphorus energy metabolism. Our results show a significant imbalance in the metabolic homeostasis during stimulation of 3T3 cells with forskolin.
Flögel
U
U
Institut für Organische Chemie, Universität Bremen, Germany.
Leibfritz
D
D
D016475:3T3 Cells; D000255:Adenosine Triphosphate / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001485:Basement Membrane / Q000737:chemistry; D005576:Colforsin / Q000494:pharmacology*; D000242:Cyclic AMP / Q000378:metabolism; D006019:Glycolysis / Q000187:drug effects; D006706:Homeostasis / Q000187:drug effects; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D010472:Perchlorates; D010767:Phosphorylcholine / Q000378:metabolism; D013268:Stimulation, Chemical
D010472:Perchlorates; D010767:Phosphorylcholine; D005576:Colforsin; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D000262:Adenylyl Cyclases