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Molecular Cloning, Genetic Mapping, and Developmental Expression of Bovine POU5F1¹

^a Hubrecht Laboratory, Netherlands Institute for Developmental Biology (NIOB), 3584 CT Utrecht, The Netherlands ^b Istituto di Anatomia degli Animali Domestici, Milan, Italy ^c Department of Herd Health and Reproduction, Faculty of Veterinary Medicine, University of Utrecht, 3584 CL Utrecht, The Netherlands ^d Laboratory of Immunogenetics, Department of Animal Sciences, University of Illinois at Urbana-Champaign, Champaign, Illinois 61820 ^e Laboratorio di Tecnologie della Riproduzione Consorzio per l'Incremento Zootecnico dell'Associazione Italiana Allevatori, 26100 Cremona, Italy ^f Virologie Immunologie Moléculaires, Institut National de la Recherche Agronomiques, 78352 Jouy-en-Josas, France ^g Institute of Reproduction and Development, Monash University, Monash Medical Centre, Clayton, Victoria, Australia

We describe isolation and characterization of the bovine ortholog of POU5F1 (bPOU5F1) encoding octamer-binding transcription factor-4 (Oct-4). The organization of bPOU5F1 is similar to its human and murine orthologs, and it shares 90.6% and 81.7% overall identity at the protein level, respectively. Transient transfection of luciferase reporter constructs in murine P19 embryonal carcinoma cells demonstrated that bPOU5F1 has a functional promoter and contains two enhancer elements, of which one is repressed by retinoic acid. bPOU5F1 was mapped to the major histocompatibility complex on chromosome 23. bPOU5F1 mRNA was detected by nested reverse transcription-polymerase chain reaction in immature oocytes and in in vitro-produced preattachment-stage embryos. Oct-4 in oocytes and in vitro-produced preattachment-stage embryos was demonstrated by indirect immunofluorescence. Confocal laser scanning microscopy revealed Oct-4 in both the inner cell mass and trophoblast cells of the blastocyst until Day 10 of development. Immunofluorescence performed on the outgrowths formed at Day 13 postfertilization from in vitro-produced Day 8 blastocysts showed Oct-4 staining in all cells. This expression pattern suggests that bPOU5F1 acts early in bovine embryonic development but that its expression is not restricted to pluripotent cells of the blastocyst.

¹ M.J.T. van Eijk was financed by the European Community's Biotechnology Program, project number EC-BIO2-CT90–0358. This work was also funded by EC-BIO2-CT92–0067 and CNR Target Project on Biotechnology.

² Correspondence: F. Gandolfi, Istituto di Anatomia degli Animali Domestici, Via D. Trentacoste 2, Milan, Italy. FAX 3902 214 0745; fulvio.gandolfi{at}unimi.it

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