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Pluripotent Lineage Definition in Bovine Embryos by Oct4 Transcript Localization¹

Center for Animal Transgenesis and Germ Cell Research, University of Pennsylvania, Kennett Square, Pennsylvania 19348

The POU-domain transcription factor Pou5f1 (Oct4) is restricted to pluripotent embryonic cells and the germ line of the mouse and is required for the maintenance of pluripotency of cells within the inner cell mass of the mouse blastocyst. Despite highly conserved genomic organization and regulatory regions between the mouse Oct4 gene and its bovine orthologue, bovine Oct4 protein is not restricted to the inner cell mass of blastocyst-stage embryos, suggesting that Oct4 may not be a key regulator of pluripotency in the bovine. We analyze the temporal and spatial distribution of Oct4 transcript in bovine oocytes and preimplantation-stage embryos, and in contrast to protein distribution, we find strong conservation between bovine and mouse. Oct4 transcript is present at low levels in the bovine oocyte. Similar to mouse, bovine Oct4 transcription begins one to two cell cycles after zygotic genome activation, followed by a sharp increase in transcription subsequent to compaction. Oct4 transcript is ubiquitously present in all cells of embryos at the morula stage; however, in Day 7 bovine blastocysts, Oct4 signal is not visible in the trophectoderm by in situ hybridization, indicating that transcriptional downregulation of Oct4 on differentiation is similar to that observed in mouse and other mammals. These results indicate that in contrast to protein distribution, regulation of Oct4 transcription is conserved between mammalian species.

¹ Supported in part by the Marion Dilley and David George Jones Funds and the Commonwealth and General Assembly of Pennsylvania (to K.J.M., S.E. and S.K.), NIH funding R01-HD-44066-01A1 (to K.J.M.), and a grant from the Lalor Foundation (to S.K. with K.J.M.).

² Correspondence: K. John McLaughlin, Center for Animal Transgenesis and Germ Cell Research, University of Pennsylvania, Kennett Square, PA 19348. FAX: 610 925 8121; kjmclaug{at}vet.upenn.edu

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