Expression of mRNAs for DNA methyltransferases and methyl-CpG-binding proteins in the human female germ line, preimplantation embryos, and embryonic stem cells

• Published in: Molecular reproduction and development Vol. 67; no. 3; pp. 323 - 336
• Main Authors: Huntriss, J., Hinkins, M., Oliver, B., Harris, S.E., Beazley, J.C., Rutherford, A.J., Gosden, R.G., Lanzendorf, S.E., Picton, H.M.
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.03.2004; Wiley-Liss
• Subjects: Biological and medical sciences; Blastocyst - metabolism; DNA methyltransferase; DNA Modification Methylases - biosynthesis; DNA Modification Methylases - genetics; DNA-Binding Proteins - biosynthesis; DNA-Binding Proteins - genetics; Embryology: invertebrates and vertebrates. Teratology; embryonic stem cell; Female; Fundamental and applied biological sciences. Psychology; genomic imprinting; Humans; inner cell mass; methyl binding domain; Molecular embryology; oocyte; Ovum - metabolism; Pregnancy; preimplantation embryo; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; Stem Cells - metabolism; Human; Embryonic development; Enzyme; Transferases; Stem cell; Germinal cell; Gene expression; In vitro; Genomic imprinting; Ovary; In vivo; Oogenesis; Methyltransferases; DNA; Female genital system; Blastocyte; Ovarian follicle; Oocyte
• ISSN: 1040-452X; 1098-2795
• DOI: 10.1002/mrd.20030

Recent evidence indicates that mammalian gametogenesis and preimplantation development may be adversely affected by both assisted reproductive and stem cell technologies. Thus, a better understanding of the developmental regulation of the underlying epigenetic processes that include DNA methylation is required. We have, therefore, monitored the expression, by PCR, of the mRNAs of DNA methyltransferases (DNMTs), methyl‐CpG‐binding domain proteins (MBDs), and CpG binding protein (CGBP) in a developmental series of amplified cDNA samples derived from staged human ovarian follicles, oocytes, preimplantation embryos, human embryonic stem (hES) cells and in similar murine cDNA samples. Transcripts of these genes were detected in human ovarian follicles (DNMT3A, DNMT3b1, DNMT3b4, DNMT1, MDBs1–4, MeCP2, CGBP), germinal vesicle (GV) oocytes (DNMT3A, DNMT3b1, DNMT1, MDBs1–4, MeCP2, CGBP), mature oocytes (DNMT3A, DNMT3b1, DNMT1, CGBP), and preimplantation embryos (DNMT3A, DNMT3b1, DNMT1, DNMT3L, MBD2, MDB4, CGBP). Differential expression of DNMT3B gene transcripts in undifferentiated (DNMT3b1) and in vitro differentiated human ES cells (DNMT3b3) further demonstrated an association of the DNMT3b1 transcript variant with totipotent and pluripotent human cells. Significantly, whilst the murine Dnmt3L gene is both expressed and essential for imprint establishment during murine oogenesis, transcripts of the human DNMT3L gene were only detected after fertilisation. Therefore, the mechanisms and/or the timing of imprint establishment may differ in humans. Mol. Reprod. Dev. 67: 323–336, 2004. © 2004 Wiley‐Liss, Inc.