1. A modified radioimmunoassay for cyclic AMP was developed from the method of Steiner et al. (1969). Cyclic [3H]AMP was used as the radioactive tracer. Free and antibody-bound nucleotides were separated by adsorption of protein to Millipore filters. The assay was used to measure amounts of cyclic AMP down to 0.1pmol in 50μl. 2. The effect of glucagon on cyclic AMP content in pieces of mature rat liver maintained for 6 days in organ culture was studied. 3. Cyclic AMP content in the tissue reached a maximum in 5–15min and then decreased. This may have been partly due to an inhibitor of glucagon action formed in the tissue. Small amounts of cyclic AMP were released into the incubation medium. 4. The maximal increase in cyclic AMP content produced by glucagon decreased over 6 days in culture. However, liver pieces cultured for 2 and 6 days were more sensitive to low concentrations of glucagon than were fresh liver pieces. Glucagon concentrations for half-maximal effects were approx. 1μm and 0.05μm for fresh liver and 2-day cultured liver respectively. 5. Insulin (3.5μm) lowered the cyclic AMP content by 30% in the presence of a submaximal glucagon concentration in liver cultured for 2 days. No effect of insulin was demonstrated on fresh liver pieces. 6. Insulin and glucagon were rapidly destroyed by fresh liver pieces.
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Research Article|
April 01 1973
The effects of glucagon and insulin on adenosine 3′:5′-cyclic monophosphate concentrations in an organ culture of mature rat liver
Kenneth Siddle;
Kenneth Siddle
1Department of Chemical Pathology, The Welsh National School of Medicine, Heath Park, Cardiff CF4 4XN, U.K.
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Barbara Kane-Maguire;
Barbara Kane-Maguire
1Department of Chemical Pathology, The Welsh National School of Medicine, Heath Park, Cardiff CF4 4XN, U.K.
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Anthony K. Campbell
Anthony K. Campbell
1Department of Chemical Pathology, The Welsh National School of Medicine, Heath Park, Cardiff CF4 4XN, U.K.
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Publisher: Portland Press Ltd
© 1973 The Biochemical Society
1973
Biochem J (1973) 132 (4): 765–773.
Citation
Kenneth Siddle, Barbara Kane-Maguire, Anthony K. Campbell; The effects of glucagon and insulin on adenosine 3′:5′-cyclic monophosphate concentrations in an organ culture of mature rat liver. Biochem J 1 April 1973; 132 (4): 765–773. doi: https://doi.org/10.1042/bj1320765
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