Production of [3H]choline-labelled metabolites from endogenously 3H-labelled phosphatidylcholine in mouse pancreatic islets.
The involvement of phosphatidylcholine (PC) hydrolysis in the regulation of insulin secretion was studied in mouse pancreatic islets prelabelled with [3H]choline. Phospholipase C (PLC) and phospholipase D (PLD) activities were demonstrated and also that of an enzyme that removes both fatty acids from PC and thus catalyses the production of [3H]glycerophosphorylcholine (GroPCho). After 2 min of incubation with 20 mM glucose a 35% increase in the content of [3H]GroPCho was observed in prelabelled islets, whereas the amount of [3H]lysoPC, [3H]phosphorylcholine (PCho) and [3H]choline was unaffected. After 30 min of incubation with 20 mM glucose, 0.2 mM tolbutamide, 40 mM KC1, 10 mM succinic acid monomethyl ester (SME) or 10 mM NaF, a 25-50% increase in [3H]GroPCho was observed. In the presence of 100 microM diazoxide or 35 microM RHC 80267 the glucose activation was attenuated. PLC was stimulated slightly by tolbutamide and 100 microM isoprenaline (isoproterenol), whereas SME decreased the amount of [3H]PCho by 10%. [3H]Choline content was increased by 25-40% in the presence of 0.16 microM 12-O-tetradecanoylphorbol 13-acetate (TPA), 10 mM NaF or 100 microM carbachol. This effect of fluoride was potentiated in the presence of 20 mM glucose. It is concluded that metabolism of PC to GroPCho may be involved in the regulation of glucose-stimulated insulin secretion, and that PLD may participate in insulin secretion evoked by TPA, carbachol and fluoride.
Capito
K
K
Department of Medical Biochemistry and Genetics, Panum Institute, University of Copenhagen, Denmark.
Hansen
S E
SE
Thams
P
P
D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured; D002794:Choline / Q000378:metabolism*; D003512:Cyclohexanones / Q000494:pharmacology; D003981:Diazoxide / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology; D005997:Glycerylphosphorylcholine / Q000378:metabolism; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D007545:Isoproterenol / Q000494:pharmacology; D007700:Kinetics; D008244:Lysophosphatidylcholines / Q000378:metabolism; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D010713:Phosphatidylcholines / Q000378:metabolism*; D011189:Potassium Chloride / Q000494:pharmacology; D011865:Radioisotope Dilution Technique; D051381:Rats; D012969:Sodium Fluoride / Q000494:pharmacology; D013386:Succinates / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014044:Tolbutamide / Q000494:pharmacology; D014316:Tritium
D003512:Cyclohexanones; D007328:Insulin; D008244:Lysophosphatidylcholines; D010713:Phosphatidylcholines; D013386:Succinates; D014316:Tritium; D005997:Glycerylphosphorylcholine; D011189:Potassium Chloride; C036535:1,6-bis(cyclohexyloximinocarbonyl)hexane; D002217:Carbachol; D012969:Sodium Fluoride; D014044:Tolbutamide; D005947:Glucose; D007545:Isoproterenol; D002794:Choline; D013755:Tetradecanoylphorbol Acetate; D003981:Diazoxide; C041105:monomethyl succinate