title: Inhibitory effect of the phosphoinositide 3-kinase inhibitor LY294002 on muscarinic acetylcholine receptor-induced calcium entry in PC12h cells.
abstract: Phosphoinositide-3 kinase (PI3K) and phospholipase C (PLC) utilize the same phosphoinositides as substrates to produce different signaling molecules. These enzymes are activated by a similar set of cell signaling mechanisms, i.e., tyrosine kinases and G proteins, and affect common cell functions, including proliferation, motility, and intracellular trafficking. Despite these similarities, the interplay between these enzymes is not well understood. To address this issue, the effects of the PI3K inhibitor LY294002 on carbachol-induced calcium increase in PC12h cells were examined. As carbachol stimulates both Gq- and Gi-coupled muscarinic acetylcholine receptors (mAChRs), PI3K and PLC are activated simultaneously in this protocol. LY294002 was found to reduce the carbachol-induced calcium increase, and the reduction was attributed to suppression of calcium entry. As LY294002 did not affect either carbachol-induced calcium release or calcium entry induced by calcium store depletion, this agent was found to suppress calcium entry directly activated by mAChRs. Although PI3K was supposed to compete for substrates with PLC, the PI3K inhibitor did not enhance PLC-dependent cellular responses. As LY294002 was still effective by treating cells after carbachol stimulation, it is likely that this agent blocks the calcium entry channels directly.
authors.affiliation: Laboratory of Cellular Neurobiology, School of Life Science, Tokyo University of Pharmacy and Life Science, 192-0392 Tokyo, Japan. moritam@ls.toyaku.ac.jp
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Morita
authors.forename: Kotaro
authors.initials: K
authors.lastname: Yoshizaki
authors.forename: Akira
authors.initials: A
authors.lastname: Nakane
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Kudo
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002217:Carbachol / Q000494:pharmacology; D002867:Chromones / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D009025:Morpholines / Q000494:pharmacology*; D018727:Muscarinic Antagonists / Q000494:pharmacology*; D016716:PC12 Cells; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051381:Rats; D010738:Type C Phospholipases / Q000502:physiology
chemical_list: D002118:Calcium; D002217:Carbachol; D002867:Chromones; D004791:Enzyme Inhibitors; D009025:Morpholines; D018727:Muscarinic Antagonists; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010738:Type C Phospholipases; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one