Sources of amino acids for protein synthesis during early organogenesis in the rat .3. Methionine incorporation

Rat conceptuses at 9.5 days post-conception were cultured for 27 h in whole rat serum. [H-3]Methionine, or rat serum proteins containing [H-3]methionine, was introduced at 24 or 6 h before the termination of the culture. The total clearance of radioactivity into the embryo and the visceral yolk sac from the two sources was measured; also the extent to which the accumulated radioactivity was acid-insoluble. Similar experiments, but using [H-3]leucine, were performed for comparison. The results indicate that free amino acid and protein can both serve as sources of amino acids for incorporation into the embryo and yolk sac, and it is estimated that in vivo over 95 per cent of the methionine (and the leucine) incorporated into these tissues arises from protein captured and digested by the polk sac. Almost all the leucine accumulated into the conceptus is present as protein, but a larger fraction of the methionine accumulated is found in acid-soluble form. When the amino acids were delivered in the form of plasma proteins, the incorporation of methionine was two to three times more efficient than that of leucine, an observation most readily explained by leucine being provided in excess of requirements. In the light of reports that an adequate concentration of free methionine is important for the normal development of rat embryos in vitro, it is concluded that, although most of the amino acid required by the embryo is supplied as protein, the small fraction supplied as free amino acid may be critical for methionine but probably not for leucine. (C) 1996 W. B. Saunders Company Ltd