Biol Reprod Keystone Symposia Conference on Frontiers in Reproductive Biology & Regulation of Fertility.
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BOR - Papers in Press, published online ahead of print May 26, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.028639
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BIOLOGY OF REPRODUCTION 71, 1108–1119 (2004)
DOI: 10.1095/biolreprod.104.028639
© 2004 by the Society for the Study of Reproduction, Inc.

Embryo

Oxygen-Regulated Gene Expression in Bovine Blastocysts1

A.J. Harvey2,3, K.L. Kind3, M. Pantaleon4, D.T. Armstrong3, and J.G. Thompson3

Research Centre for Reproductive Health,3 Department of Obstetrics and Gynaecology, The University of Adelaide, The Queen Elizabeth Hospital, Woodville, South Australia 5011, Australia Department of Physiology and Pharmacology,4 University of Queensland, St. Lucia, Queensland 4072, Australia

Oxygen concentrations used during in vitro embryo culture can influence embryo development, cell numbers, and gene expression. Here we propose that the preimplantation bovine embryo possesses a molecular mechanism for the detection of, and response to, oxygen, mediated by a family of basic helix-loop-helix transcription factors, the hypoxia-inducible factors (HIFs). Day 5 compacting bovine embryos were cultured under different oxygen tensions (2%, 7%, 20%) and the effect on the expression of oxygen-regulated genes, development, and cell number allocation and HIF{alpha} protein localization were examined. Bovine in vitro-produced embryos responded to variations in oxygen concentration by altering gene expression. GLUT1 expression was higher following 2% oxygen culture compared with 7% and 20% cultured blastocysts. HIF mRNA expression (HIF1{alpha}, HIF2{alpha}) was unaltered by oxygen concentration. HIF2{alpha} protein was predominantly localized to the nucleus of blastocysts. In contrast, HIF1{alpha} protein was undetectable at any oxygen concentration or in the presence of the HIF protein stabilizer desferrioxamine (DFO), despite being detectable in cumulus cells following normal maturation conditions, acute anoxic culture, or in the presence of DFO. Oxygen concentration also significantly altered inner cell mass cell proportions at the blastocyst stage. These results suggest that oxygen can influence gene expression in the bovine embryo during postcompaction development and that these effects may be mediated by HIF2{alpha}.

1 Supported by a National Health and Medical Research Council Project Grant (157941) and by a TQEH Research Foundation Postgraduate Research Scholarship to A.J.H.

2 Correspondence. FAX: 61 8 82227521; alexandra.harvey{at}adelaide.edu.au






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Copyright © 2004 by the Society for the Study of Reproduction.