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Human Molecular Genetics Advance Access originally published online on May 18, 2004
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Human Molecular Genetics, 2004, Vol. 13, No. 14 1461-1470
DOI: 10.1093/hmg/ddh157
Human Molecular Genetics, Vol. 13, No. 14 © Oxford University Press 2004; all rights reserved

The unique transcriptome through day 3 of human preimplantation development

Anthony T. Dobson1,2,*, Rajiv Raja3, Michael J. Abeyta1, Theresa Taylor3, Shehua Shen1,2, Christopher Haqq4 and Renee A. Reijo Pera1

1Center for Reproductive Sciences, Department of Obstetrics, Gynecology and Reproductive Sciences, Departments of Physiology and Urology, Programs in Human Genetics, Cancer Genetics, and Development and Stem Cell Biology, University of California at San Francisco, San Francisco, CA 94143, USA, 2Center for Reproductive Health, University of California at San Francisco, 2356 Sutter Street, San Francisco, CA 94115, USA, 3Arcturus Bioscience Inc., 400 Logue Avenue, Mountain View, CA 94043, USA and 4Comprehensive Cancer Center, Prostate Cancer Program, University of California at San Francisco, San Francisco, CA 94143, USA

Received March 9, 2004; Accepted May 11, 2004

Successful human development is dependent upon a cascade of events following fertilization. Unfortunately, knowledge of these critical events in humans is remarkably incomplete. Although hundreds of thousands of human embryos are cultured yearly at infertility centers worldwide, the vast majority fail to develop in culture or following transfer to the uterus. In this study, we sought to characterize global patterns of gene expression in individual, normal embryos during the first three days of embryonic life using microarrays; we then compared gene expression between normally growing and growth-arrested embryos using quantitative PCR. Our results documented several novel findings. First, we found that a complex pattern of gene expression exists; most genes that are transcriptionally modulated during the first three days following fertilization are not upregulated, as was previously thought, but are downregulated. Second, we observed that the majority of genes exhibiting differential expression during preimplantation development are of unknown identity and/or function. Third, we show that embryonic transcriptional programs are clearly established by day 3 following fertilization, even in embryos that arrested prematurely with 2-, 3- or 4-cells. This indicates that failure to activate transcription is not associated with the majority of human preimplantation embryo loss. Finally, taken together, these results provide the first global analysis of the human preimplantation embryo transcriptome, and demonstrate that RNA can be amplified from single oocytes and embryos for analysis by cDNA microarray technology, thus lending credence to additional studies of genetic regulation in these cell types, as well as in other small biological samples.

* To whom correspondence should be addressed at: Center for Reproductive Sciences, 513 Parnassus Avenue, Box 0556, University of California at San Francisco, San Francisco, CA 94143-0556, USA. Tel: +1 4155140698; Fax: +1 4154763121; Email: adobson@itsa.ucsf.edu


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