Transport and uptake of retinol during chicken oocyte growth

Most, if not all, components found in the yolk of a chicken egg are extracted from the plasma compartment during the rapid growth phase of the oocyte. Uptake of the major yolk constituents, very-low-density lipoprotein and vitellogenin, has been shown to be mediated by a specific receptor in the plasma membrane of the oocyte (Barber, D.L., Aebersold, R., Sanders, E.J. and Schneider, W.J. (1991) J. Biol. Chem. 266, 18761-18770). In the current study, we sought biochemical evidence for the uptake into oocytes of a minor but biologically very important component, the vitamin retinol. For transport in serum, retinol is bound to retinol-binding protein (RBP), which in turn forms a complex with transthyretin (TTR). In order to gain insight into the biochemical details of transport of the vitamin, we have identified, purified and characterized RBP, TTR, and RBP-TTR complexes from chicken serum and yolk. The results demonstrate that both serum and yolk contain the tertiary retinol-RBP-TTR complexes as well as free RBP and TTR. Western blots of yolk collected from oocytes at different stages of growth show that both RBP and TTR, but not albumin, are more abundant at early stages relative to total yolk protein. In addition, we find both RBP and TTR in endocytic clathrin-coated vesicles of the oocyte. Our results support the hypothesis that retinol, which must be imported by the oocyte for proper embryonic development, is internalized by the chicken oocyte bound to its serum protein-transport complex.