A protein-binding radioassay for cyclic AMP was modified to detect less than 0.025pmol of the nucleotide. The method was applied to the measurement of cyclic AMP in small numbers of mouse pancreatic islets (as little as 25μg of tissue) by use of barium acetate–H2SO4 for deproteinization. The concentration of cyclic AMP in mouse islets incubated in media containing 3.3 or 20mm-glucose was 0.016pmol/10 islets (approx. 1μm in intracellular water). Glucose concentration (3.3 or 20mm) had no detectable effect on islet concentrations of cyclic AMP with periods of incubation or perifusion ranging from 0.5 to 60min, although insulin release rate was rapidly increased by 20mm-glucose. Caffeine (5mm) or 3-isobutyl-1-methylxanthine (1mm), which are known inhibitors of islet cyclic AMP phosphodiesterase, produced marked and rapid increases in islet cyclic AMP concentration at 3.3 or 20mm-glucose, but only enhanced the insulin release rate at the higher glucose concentration. The role of cyclic AMP in insulin release induced by glucose is discussed.
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June 1973
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Research Article|
June 01 1973
Concentration of adenosine 3′:5′-cyclic monophosphate in mouse pancreatic islets measured by a protein-binding radioassay Available to Purchase
R. H. Cooper;
R. H. Cooper
1Department of Biochemistry, University of Bristol, Bristol BS8 1TD, U.K.
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S. J. H. Ashcroft;
S. J. H. Ashcroft
1Department of Biochemistry, University of Bristol, Bristol BS8 1TD, U.K.
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P. J. Randle
P. J. Randle
1Department of Biochemistry, University of Bristol, Bristol BS8 1TD, U.K.
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Publisher: Portland Press Ltd
© 1973 London: The Biochemical Society
1973
Biochem J (1973) 134 (2): 599–605.
Citation
R. H. Cooper, S. J. H. Ashcroft, P. J. Randle; Concentration of adenosine 3′:5′-cyclic monophosphate in mouse pancreatic islets measured by a protein-binding radioassay. Biochem J 1 June 1973; 134 (2): 599–605. doi: https://doi.org/10.1042/bj1340599
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