Protein synthesis in mature human oocytes.
As a first step toward understanding control of gene expression in early human development, an analysis of protein synthesis and amino-acid transport in unfertilized mature oocytes was initiated. Qualitative patterns of protein synthesis were examined in individual oocytes cultured in medium containing radiolabeled methionine. No differences in synthetic pattern of proteins, resolved by one-dimensional electrophoresis and fluorography, were observed in oocytes analyzed from times varying from 12 to 52 hr following collection by laparoscopy. Contamination by follicular or corona radiata cells was readily distinguished on the basis of increased relative synthesis of a polypeptide with Mr = 44,000, a dominant product of synthesis in follicular cells. Based on the specific activity of the methionine precursor, the absolute rate of synthesis was measured to be about 50 pg/oocyte/hr, a value higher than in mouse unfertilized eggs. No difference in protein synthetic rate was observed in oocytes analyzed at 12 hr postcollection versus later times up to 50 hr postcollection. Competition of methionine uptake by leucine, efflux of radiolabeled methionine from preloaded oocytes into medium containing methionine and uptake of methionine in medium with low sodium ion concentration was observed. These findings are consistent with the presence of an L (leucine-preferring) system for neutral amino acid transport, similar to that in mouse and rabbit eggs. Total protein was measured to be about 150 ng/oocyte, a value five times that of the mouse. These studies provide basic data for further analysis of oocytes and perhaps preimplantation stage embryos in the future.
D000818:Animals; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D014176:Protein Biosynthesis; D011506:Proteins / Q000302:isolation & purification
D011506:Proteins; D008715:Methionine