Insulin action in intact mouse diaphragm I.

Summary The incubation of intact mouse diaphragms with insulin caused a dose and time dependent increase in the independent activity of glycogen synthase in tissue extracts. 2-deoxyglucose (2–10 mm) alone markedly stimulated the conversion of glycogen synthase to the independent activity under conditions in which tissue ATP concentrations were not affected. The incubation of diaphragms with both insulin and 2-deoxyglucose resulted in a greater than additive effect. Insulin stimulated the uptake of 2-deoxyglucose into mouse diaphragms, accumulating as 2-deoxyglucose-6-phosphate. The accumulation of 2-deoxyglucose-6-phosphate correlated well with the increase in the independent activity of glycogen synthase and with the activation of glycogen synthase phosphatase in tissue extracts. The uptake of 3-0-methyl glucose was also markedly stimulated by insulin, without affecting the activity of glycogen synthase. Both glucose-6-phosphate and 2-deoxyglucose-6-phosphate stimulated the activation of endogenous glycogen synthase phosphatase activity in muscle homogenates. We conclude that insulin, in addition to its effects in the absence of exogenous sugars, increases the independent activity of glycogen synthase through increased sugar transport resulting in increased concentrations of sugar-phosphates which promote the activity of glycogen synthase phosphatase.Abbreviations GS Glycogen synthase - GS-I Glycogen synthase activity independent of G6P - GS-D Glycogen synthase activity dependent on G6P - G6P Glucose-6-phosphate - ATP Adenosine triphosphate - EDTA Ethylene diamine tetracetic acid - Mops Morpholinopropane sulfonic acid - 2DG 2-Deoxy glucose - 3-0-MG 3-0-Methyl glucose - tricine N-tris(Hydroxymethyl)methyl glycine Enzymes: Glycogen Synthase — UDPGlucose — Glycogen Glucosyl — Transferase (EC 2.4.1.11) J. Larner is an established investigator of the American Diabetes Association.