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Hum. Reprod. Advance Access first published online on February 10, 2005
This version published online on March 10, 2005
Human Reproduction, doi:10.1093/humrep/deh778
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Human Reproduction © European Society of Human Reproduction and Embryology 2005; all rights reserved
Received September 7, 2004
Accepted January 12, 2005

Article

Expression of genes regulating chromosome segregation, the cell cycle and apoptosis during human preimplantation development

D. Wells 1*, M.G. Bermudez 2, N. Steuerwald 3, A.R. Thornhill 4, D.L. Walker 4, H. Malter 5, J.D.A. Delhanty 6, and J. Cohen 5

1 Department of Obstetrics & Gynecology, Yale University Medical School, 333 Cedar Street, New Haven, CT 06520, Reprogenetics LCC, 101 Old Short Hills Road, Suite 501, West Orange, NJ 07052, Tyho-Galileo Research Laboratories LLC, 101 Old Short Hills Rd, Suite 501, West Orange, NJ 07052
2 Reprogenetics LCC, 101 Old Short Hills Road, Suite 501, West Orange, NJ 07052
3 Reprogenetics LCC, 101 Old Short Hills Road, Suite 501, West Orange, NJ 07052, Tyho-Galileo Research Laboratories LLC, 101 Old Short Hills Rd, Suite 501, West Orange, NJ 07052
4 Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Mayo Clinic, Rochester, Minnesota 55905 and
5 Tyho-Galileo Research Laboratories LLC, 101 Old Short Hills Rd, Suite 501, West Orange, NJ 07052
6 Department of Obstetrics and Gynaecology, University College London, 86-96 Chenies Mews, London WC1E 6HX, UK

* To whom correspondence should be addressed.
D. Wells, E-mail: dagan.wells{at}yale.edu


  Abstract

BACKGROUND: Appropriate gene expression is vital for the regulation of developmental processes. Despite this fact there is a remarkable paucity of information concerning gene activity during preimplantation development. METHODS: We employed reverse transcription and real-time fluorescent PCR to quantify the expression of nine genes (BRCA1, BRCA2, ATM, TP53, RB1, MAD2, BUB1, APC and {beta}-actin) in oocytes and embryos. A full characterization of all genes was achieved in 42 embryos and four oocytes. The genes analysed have a variety of important cellular functions. RESULTS: Oocytes displayed relatively high levels of mRNA transcripts, while 2-3-cell embryos were seen to contain very little mRNA from any of the genes examined. Recovery of expression levels was not seen until the 4-cell stage or later, with the presumptive activation of the embryonic genome. Some genes displayed sharp increases in expression in embryos composed of 4-8 cells, but, for most, maximum expression was not achieved until the blastocyst stage. CONCLUSIONS: Our data show that it is possible to define characteristic gene expression profiles for each stage of human preimplantation development. The identification of genes active at defined preimplantation phases may provide clues to the cellular pathways utilized at specific stages of development. Expression of genes that function in DNA repair pathways indicate that DNA damage may be common at the cleavage stage. We suggest that specific patterns of gene expression may be indicative of embryo implantation potential.

Keywords: apoptosis; cell cycle checkpoint; IVF; PGD; RT-PCR.
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