Synthesis of Retinol-binding Protein and Transthyretin in Yolk Sac and Fetus in the Rat

Abstract

Levels of retinol-binding protein (RBP) and transthyretin (TTR) were determined in rat maternal livers, placenta, yolk sac, whole fetuses and fetal livers at different stages of gestation. Yolk sac concentrations of RBP and TTR expressed as micrograms per mg protein were three- to fivefold higher than liver values. TTR (moles) in the whole fetus was higher than RBP at all stages of gestation. Fetal hepatic RBP concentration was relatively constant throughout gestation, where fetal hepatic TTR concentration was low until close to parturition. RBP was observed in fetal microsomes at 12 d gestation. Incorporation of labeled amino acids into both RBP and TTR in vitro was observed in the yolk sac. In the 20-d fetal liver, incorporation to RBP and TTR was observed, whereas at 14 d gestation, incorporation only to RBP was observed. A small amount of synthesis of both proteins was also observed in the placenta. In the fetal circulation at 20–21 d gestation, no TTR-RBP complex was observed; instead a broad peak of RBP was found eluting at 20,000–40,000 daltons on gel chromatography. Incubation of fetal serum with maternal TTR resulted in an RBP peak eluting with an Mr of approximately 40,000. Treatment of the fetal serum with either lysozyme, neuraminidase or endoglycosidase H resulted in a 20,000 dalton RBP peak and following incubation with maternal TTR, a 70,000 dalton RBP-TTR complex was formed. Yolk sac, fetal liver and amniotic fluid on gel filtration exhibited a 40,000–20,000 dalton RBP peak. It is suggested that retinol transport in the fetus may involve RBP and TTR synthesized in the yolk sac as well as in fetal tissue. Results obtained after glycosidase treatment suggest that RBP or TTR in the fetus may be glycosylated and thus differ from the adult proteins.