Developmental increases in glucose transporter concentration in the sheep placenta

To explore the molecular basis for the gestational increase in glucose transport capacity of the sheep placenta in vivo, placentas from twin-pregnant ewes at days 75, 110, and 140 postcoitus (n = 6/group) were analyzed for glucose transporter (GT) concentration. Concentration (pmol/mg protein) of D-glucose-inhibitable binding sites, measured by [H-3]cytochalasin B binding analysis, increased 3.4 times from mid-to late pregnancy. Concurrently abundance of GLUT-1 and GLUT-3 protein, measured by immunoblotting with specific polyclonal antibodies, increased 2.3 and 2.9 times, respectively, while abundance of GLUT-1 and GLUT-3 mRNA, measured by Northern blotting, increased 1.8 and 5.9 times, respectively. GLUT-4 protein was undetectable at all stages of pregnancy. Quantitative immunoblotting-indicated that GLUT-1 accounted for 86.8 +/- 1.6% at day 75 and 56.1 +/- 4.1% at day 140 of total cytochalasin B binding sites. Thus increases in GT concentration explain much of the gestational increase in glucose transfer capacity observed in vivo. The gestational decline in relative contribution of GLUT-1 to cytochalasin binding, together with the greater developmental increases in GLUT-3 mRNA and protein, further suggests that the relative importance of GLUT-3 increases with gestational age.