Insulin-sensitive glucose transporter transcript levels in calf muscles assessed with a bovine GLUT4 cDNA fragment

Abstract

Previous studies have shown that the expression of the insulin-sensitive glucose transporter (GLUT4) is lower in oxidative muscles than in glycolytic muscles in bovines and goats in contrast to observations in rats. Additional experiments in this work provide very strong arguments that the immunoreactive band detected does represent GLUT4 protein, which further validates our previous results. Therefore, to determine the level of regulation, the main objective of the present study was to measure GLUT4 mRNA amounts in various bovine muscles. A 241-bp fragment of the bovine GLUT4 cDNA was cloned by polymerase chain reaction (PCR). It shares 80–90% sequence identity with related sequences in other species. This PCR-amplified bovine GLUT4 probe was used to determine the distribution of GLUT4 mRNA in bovine tissues in comparison with that of GLUT1 mRNA. Moreover, GLUT4 mRNA amounts were quantified by Northernblot analysis in heart and seven skeletal muscles with various oxidative and glycolytic activities from seven ruminant calves. GLUT4 mRNA was detected by Northern-blot analysis only in calf insulin-sensitive tissues. In contrast, GLUT1 mRNA was detected in all tissues studied except liver. GLUT4 mRNA amount was the highest in masseter and heart, which are oxidative muscles (1.67 ± 0.16 and 1.53 ± 0.19 units/g wet tissue weight, respectively) and the lowest in glycolytic or oxido-glycolytic muscles (0.31 ± 0.04 to 1.00 ± 0.09 units/g wet tissue weight; SEM, n = 7). These data and our previous results provide evidence for translational and/or posttranslational control mechanisms of bovine GLUT4 protein expression in a muscle type-specific manner.