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 December 10, 2003.
Biol Reprod 2003, 10.1095/biolreprod.103.022236
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Submitted August 12, 2003
Returned for revision August 27, 2003
Accepted December 1, 2003

Embryo


Analysis of Imprinted Messenger RNA Expression During Bovine Preimplantation Development

Nancy T Ruddock *, Katrina J Wilson , Melissa A Cooney , Natasha A Korfiatis , R Tayfur Tecirlioglu , and Andrew J French

* To whom correspondence should be addressed. E-mail: nancy.ruddock{at}med.monash.edu.au.

Abstract
While the expression and epigenetic differences of imprinted genes has been extensively characterized in the mouse and human, little is known about imprinted genes in livestock species. In the current study, eight genes that are imprinted in the human or mouse, were investigated in preimplantation bovine embryos. Amplified cDNA was created from three single MII oocytes or embryos throughout preimplantation development. The imprinted genes Dlk1 and Mest (isoform1) had no detectable transcripts during preimplantation development. Gnas and Grb10 were expressed in most embryos from the 2-cell to blastocyst stages of development. Mest (isoform2) was expressed in all oocytes and embryos, except for one blastocyst sample. Ndn and Xist were expressed from the 8-16-cell stage (MZT) onwards. Sgce was expressed until the MZT, and Nnat in both early ({alpha} form) and late ({beta} form) stage embryos. The paternally imprinted genes Gnas, Grb10 and Xist were expressed in both IVF and parthenogenetic (PA) blastocysts as expected. Of the 4 maternally imprinted genes expressed in the blastocyst (Mest, Ndn, Nnat and Sgce), Nnat alone showed differential mRNA expression between IVF and PA blastocysts suggesting imprinting by this stage of development. In conclusion, 7 of the 8 genes investigated showed mRNA expression during preimplantation development indicating a potential role during early development. Also significant is the observation that Nnat is imprinted by the blastocyst stage of development while the other genes are not, indicating a temporal imprinting program.

Key words: Embryo • Developmental biology • Early development • Gene regulation • In vitro fertilization


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