The quantity and distribution of transferrin and transferrin-binding sites in the placenta were investigated in rabbits on the 28th-29th days of pregnancy. The animals were injected intravenously with a mixture of 59Fe-125I-labelled rabbit diferric transferrin and 131I-labelled rabbit albumin. The binding of transferrin to placentas removed 3-75 min later was determined by using the 131I-labelled albumin values to correct for tissue content of plasma. Mean values for transferrin binding of 1460 and 560 micrograms/g tissue were obtained 3-15 and 45-75 min after injection, respectively. Gel filtration of placental extracts prepared with the non-ionic detergent, Teric 12A9, showed that the 125I-labelled transferrin bound to a large molecular weight component which had the properties of a specific receptor. The receptor had a higher affinity for diferric transferrin than for apotransferrin. The subcellular distribution of transferrin binding sites was determined by differential centrifugation of placental homogenates and by electron microscope autoradiography. The results with the former method indicated that the transferrin was bound to the microsomal fraction of the cells. Autoradiography showed that the majority of the transferrin molecules were at intracellular sites, mainly on the membrane of intracellular vesicles. It is concluded that iron-containing transferrin molecules enter the trophoblast cells by endocytosis or via a canalicular system after binding to cell membrane receptors. The higher affinity of the receptors for diferric transferrin than for apotransferrin explains the difference in amount of transferrin binding found within 15 min of injecting labelled diferric transferrin and that found 45-75 min later when much of the iron had been removed from the transferrin.
Animals Autoradiography