I n situ hybridization was used to evaluate patterns of gene expression for glucose transporters 1-4 (GT1-4) in the rat uteroplacenta from the time of implantation through term, and in vivo regional placental glucose metabolism was measured by 14C-labeled 2-deoxyglucose uptake. GT1 mRNA was highly abundant and GT3 was barely detected in the postimplantation decidual reaction. GT1 and 3 mRNAs were colocalized in the labyrinthine syncitiotrophoblast layer of the chorioallantoic placenta, which forms the membranous barrier between maternal and fetal circulations. The level of labyrinthine GT3 mRNA showed no change from midgestation through term; however, the volume of the labyrinth and hence total GT 3 gene expression increased greatly during this period. Labyrinthine GT1 mRNA levels, in contrast, showed significant diminution near term. GT1 mRNA was also localized in the placental growth plate, or junctional zone, where it was most abundant during the period of rapid placental growth and was decreased at term. Placental glucose metabolism, as reflected by steady-state 2-deoxyglucose uptake, was highest in the junctional zone during the rapid growth phase during midgestation, and decreased significantly at term, in parallel with GT1 gene expression. These findings suggest that GT1 is responsible for supplying glucose for use as a placental fuel and that GT3 is important for glucose transfer to the embryo.