Effect of insulin on protein synthesis in skeletal muscle of an isolated perfused preparation of rat hemicorpus.

Abstract

A method for perfusion in vitro of a preparation of rat hemicorpus was developed for study of the metabolism of skeletal muscle. The preparation was stable during perfusion, as indicated by maintenance of ATP concentration, perfusion pressure, and oxygen consumption for up to 90 min. The perfused hemicorpus provided the following advantages for study of protein synthesis in skeletal muscle: (a) hormones and substrates reached the muscle cells through an intact capillary bed, and (b) the preparation included the psoas muscle, which was sufficiently large to allow measurements of intermediates in the pathway of protein synthesis and was readily homogenized for preparation of ribosomes and ribosomal subunits. Perfusion of psoas muscle from fasted rats with buffer containing glucose and insulin reduced the concentration of ribosomal subunits and increased phenylalanine incorporation as compared to perfusion with buffer containing glucose alone. In addition, the hormone increased glucose uptake from the perfusate and inhibited release of free fatty acids from the preparation. When the muscle was perfused with buffer that contained glucose and palmitate, the concentration of ribosomal subunits and phenylalanine incorporation were unchanged. Since fatty acid is known to stimulate protein synthesis in heart muscle, these results indicated that rates of protein synthesis in heart, but not in skeletal, muscle would be maintained during fasting or in diabetic animals by increased plasma concentration of fatty acid.