EFFECTS OF INSULIN ON THE INTACT LEVATOR ANI MUSCLE OF THE RAT

Abstract

ABSTRACT The effects of insulin on the rate of intracellular accumulation and incorporation into the protein of some normal amino acids (glycine-3H, leucine-14C and valine-14C) in the intact levator ani muscle of the rat were studied in vitro. The effect of puromycin on the accumulation of these amino acids as well as on the accumulation of the model amino acid α-aminoisobutyric acid-14C (AIB-14C) was examined. In addition the effect of insulin on the incorporation of adenine-14C into the muscle RNA as well as on the effect of actinomycin D added to the incubation medium were also investigated. The insulin stimulated AIB-14C transport is described by the formulation of Michaelis-Menten and approximate values of Km and Vmax are calculated for this transport in the levator ani muscle. Insulin was found to stimulate the intracellular accumulation of glycine-3H and AIB-14C, while a significant decrease was found in the accumulation of leucine-14C and valine-14C. Insulin also stimulated the incorporation of glycine-3H and leucine-14C into the muscle protein. When puromycin was added to the medium, no radioactivity was found in the protein fraction and insulin then stimulated the intracellular accumulation of glycine-3H, leucine-14C and AIB-14C. A slight but significant stimulation with insulin was found on the incorporation of adenine-14C into the RNA-fraction of the levator ani muscle. The addition of actinomycin D was found to inhibit this effect of insulin, but did not change the insulin stimulation of the AIB-14C accumulation. The results are discussed in relation to our present knowledge of the different in vitro effects of insulin.