Acute exercise, epinephrine, and diabetes enhance insulin binding to skeletal muscle.

We measured insulin binding to crude membranes from rat skeletal muscle, with binding expressed relative to the sarcolemmal marker, cholesterol ester (CE). The amount of CE in the sarcolemma remained constant after streptozotocin-induced diabetes and acute exercise (swam for 90 min). Soleus (predominantly slow-twitch fibers) had higher insulin binding capacity than extensor digitorum longus (predominantly fast twitch). Both diabetes and acute exercise enhanced insulin binding. The shape of the enhanced insulin binding curve differed, however, between diabetes and acute exercise. Diabetes elicited a uniform increase in binding across the insulin concentrations measured (0.04-166 nM); acute exercise elicited the largest increase at the lower concentrations, suggesting different mechanisms cause the enhanced binding. Addition of 13.1 nM epinephrine to the perfusate in a rat hindlimb preparation increased insulin binding in a pattern similar to acute exercise. In contrast, muscular contraction stimulated by the sciatic nerve (1 Hz) or reduction of perfusate insulin from 100 to 40 pM, two additional correlates of acute exercise, had no effect. The increased insulin binding after acute exercise, therefore, appears to be mediated through elevated levels of catecholamines and not upregulation of the insulin receptor.

Tumorigenicity testing of primate cell lines in nude mice, muscle organ culture and for colony formation in soft agarose.

Primate neoplastic and finite cell lines were tested in one in vivo and two in vitro test systems: adult nude mice, muscle organ culture (MOC) and soft agarose (SA). Comparison of the sensitivity of the systems indicated that nude mice were inferior to either in vitro system: WI-38 VA13 (an SV40 transformed cell line) did not cause tumours in these animals yet it behaved as if it were neoplastic in MOC and formed colonies in SA. There was complete correlation between results obtained in MOC and SA. All cell lines which produced tumors in vivo were positive in both in vitro test systems. None of the lines which showed normal patterns in MOC and in SA was tumorigenic in nude mice. Since testing in vitro is simpler, faster, and is thought to be reliable, we recommend SA followed by MOC as the initial assays for determining tumorigenicity of cells.