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  • 1
    ISSN: 1432-0428
    Keywords: Alloxan ; cyclic AMP ; isolated islets ; insulin secretion ; glucose metabolism ; 3-0-methylglucose ; glyceraldehyde
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Insulin secretion was stimulated and cyclic adenosine 3′, 5′-monophosphate (cAMP) levels were elevated in isolated rat islets by 27.5 mmol/l glucose. Alloxan caused a dose-dependent decrease in both variables with complete obliteration of insulin release at a concentration of 1.25 mmol/l. D-glucose, in the presence or absence of extracellular calcium, or 3-0-methyl-D-glucose (both at 27.5 mmol/l) protected completely against the effects of alloxan on both glucose-induced insulin release and cAMP levels. 3-0-Methylglucose did not stimulate insulin secretion or elevate cAMP and did not interfere with glucose-stimulated secretion or elevation of cAMP. When glucose-stimulated insulin release was abolished by alloxan, the metabolism of glucose, determined by the rate of3H2O formation from [5-3H] glucose, was depressed by 20%. It is concluded that alloxan altered the adenylate cyclase system such that it could no longer be stimulated by glucose. Glucose-stimulated insulin secretion or elevation of cAMP did not appear essential for glucose to protect against alloxan. Protection by 3-0-methylglucose did not appear to be mediated through an alteration of cAMP metabolism. Alloxan did not inhibit glucose-induced insulin secretion by grossly altering glycolysis.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0428
    Keywords: Glucose ; cyclic AMP ; calcium ; insulin ; insulin secretion ; receptor mechanism ; second messenger
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Adenosine 3′,5′-cyclic monophosphate (cAMP) levels of isolated perifused pancreatic islets were elevated by high levels of glucose concomitantly with initiation of enhanced insulin secretion. The rise of cAMP was biphasic and seemed to be related to the temporal biphasic kinetics of insulin release. However, the temporal profiles of cAMP level changes and of insulin release differed; the major rise of the cAMP levels was seen during the initial phase, whereas insulin secretion was more pronounced during the second phase of release. Glucose-induced cAMP elevation required the presence of extracellular Ca++. Mannoheptulose completely blocked cAMP elevation due to high glucose. Exogenous insulin which has been shown by others to inhibit insulin secretion in vitro, blunted the glucose-induced cAMP rise. These observations and data in the literature are compatible with the concept that under physiological conditions glucose governs the intracellular cAMP levels in a Ca++ dependent manner — either directly or indirectly through metabolic effects.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0428
    Keywords: Keywords MODY-2 ; glucokinase ; glucose threshold ; insulin secretion ; beta-cell ; mathematical model.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Aims/hypothesis. Mutations of the glucokinase gene cause hyperglycaemia or hypoglycaemia. A quantitative understanding of these defects of glucose homeostasis linked to the glucokinase gene was lacking. Therefore a database of kinetic variables of wild-type and 20 missense mutants of glucokinase was developed and used in mathematical modelling to predict the thresholds for glucose-stimulated insulin release. Methods. Recombinant human glucokinase was generated in E. coli. The kcat, glucose S0.5, ATP Km, and Hill number of glucokinase were determined. Inhibition by Stearoyl CoA and glucokinase regulatory protein and thermal stability were assayed for all mutants kinetically similar to wild-type glucokinase. A mathematical model predicting the threshold for glucose-stimulated insulin release was constructed. This model is based on the two substrate kinetics of glucokinase and the kinetic variables of the database. It is assumed that both glucokinase gene alleles are equally expressed in beta-cells and that induction of glucokinase occurs as a function of basal blood glucose. Results. Large changes, varying greatly between mutants were found in nearly all variables. Glucokinase flux at threshold for glucose-stimulated insulin release was about 25 % of total phosphorylating potential in the normal beta-cell and this was used to predict thresholds for the mutant heterozygotes. Clinical data for maturity onset diabetes of the young type linked to the glucokinase gene and familial hyperinsulinaemic hypoglycaemia linked to the glucokinase gene and the glucokinase kinetic data of this study were used to test the model. The model predicts fasting blood glucose between 3 and 7 mmol/l in these cases. Conclusion/interpretation. A kinetics database of wild-type and 20 mutants of glucokinase was developed. Many kinetic differences were found for the mutants. The mathematical model to calculate the threshold for glucose-stimulated insulin release predicts fasting blood glucose between 3 and 7 mmol/l in subjects with glucokinase gene mutations. [Diabetologia 42: 1175–1186]
    Type of Medium: Electronic Resource
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