Isolated cardiac myocytes. A new cellular model for studying insulin modulation of monosaccharide transport

Abstract

The usefulness of isolated Ca2+-tolerant myocytes as a cellular model system for investigating modulation of monosaccharide transport by insulin was investigated. We have found that the isolation technique described by Haworth et al. (Haworth, R.A., Hunter, D.R. and Berkoff, H.A. (1980) J. Mol. Cell. Cardiol. 12, 715-724), with some minor modifications, consistently gave the highest yield of quiescent, rod-shaped myocytes which maintained their integrity in the presence of 2 mM calcium. Using 3-0-methylglucose, a non-metabolized sugar, transport was shown to possess saturability, substrate stereospecificity, competition and countertransport; all of which have been thoroughly established for D-glucose transport in other systems. The apparent Km of transport ranged from 2.3 to 3.5 mM. Insulin (10 nM) caused a small but significant increase in Km and a 2-3-fold increase in Vmax. These results suggest that this myocyte preparation will provide a useful model for studying the transport-related effects of insulin as well as current hypotheses regarding the mechanism of insulin modulation of transport at the cellular level.