Some factors affecting the response of muscle to insulin.

1. The effect of various changes in the composition of the supporting medium on the capacity of isolated rat diaphragm to incorporate amino acids into its protein has been studied. 2. Replacement of most of the normal ionic constituents by sucrose is inhibitory towards protein synthesis, as is also substitution of choline or K(+) for Na(+). 3. The capacity of the tissue to respond to a stimulatory effect of insulin is impaired in the sucrose media and under certain conditions in the absence of Na(+), particularly when Na(+) is replaced by K(+) and the (14)C-labelled amino acid is presented at a relatively high concentration. 4. Cutting of the tissue before incubation also decreases incorporating capacity and markedly decreases responsiveness to insulin. 5. In abnormal media the cellular content of ATP falls sharply. 6. The ATP content of the tissue also declines in the presence of 2-deoxyglucose. This change is prevented by the addition of glucose but not of pyruvate and succinate. 7. Although affecting the rate of amino acid incorporation the ATP content is not thought generally to limit sensitivity to insulin.