Methylation in fertilised and parthenogenetic preimplantation mouse embryos.
DNA methylation is one of the proposed biochemical mechanisms involved in cell differentiation and in genomic imprinting, and DNA methyltransferase (DMT) is a key enzyme in the embryo since mutation of its gene is lethal early in development. In order to verify that non-viability of uniparental embryos was not due to a defect in the regulation of DMT activity, we compared the metabolism of methylation in parthenogenetic embryos (maternal genome) and in fertilised embryos (maternal and paternal genomes). As regards total methylation, estimated by a measure of S-adenosyl methionine (SAM) and S-adenosyl homocysteine (SAH) formation, no significant difference was found between the two kinds of embryos during preimplantation development. Mean values were 4.5 +/- 0.6 fmol (SAM+SAH)/h per 2-cell embryo and 0.40 +/- 0.05 fmol SAH/h per 2-cell embryo, i.e. a SAH/(SAM+SAH) ratio of 9%; there was no detectable SAH formation in blastocysts. The same observation can be made for DMT activity, with mean values of: 7.8 fmol/h per oocyte, 8.5 fmol/h per 2-cell embryo, 6.1 fmol/h per 4-cell embryo, 4.1 fmol/h per morula, and no detectable activity in blastocysts. Total methylation as well as DNA methylation is characterised by a progressive drop in activity during preimplantation development.
Croteau
S
S
Laboratoire de Biologie Appliquée, INSA, Villeurbanne, France.
Menezo
Y
Y
D000818:Animals; D015254:DNA Modification Methylases / Q000378:metabolism; D004622:Embryo, Mammalian / Q000201:enzymology / Q000378:metabolism*; D047108:Embryonic Development; D005260:Female; D005306:Fertilization; D018392:Genomic Imprinting; D006710:Homocysteine / Q000031:analogs & derivatives / Q000096:biosynthesis; D008715:Methionine / Q000378:metabolism; D008745:Methylation; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D010312:Parthenogenesis; D011247:Pregnancy; D012436:S-Adenosylmethionine / Q000096:biosynthesis
D006710:Homocysteine; D012436:S-Adenosylmethionine; D008715:Methionine; D015254:DNA Modification Methylases