Molecular cloning, genetic mapping, and developmental expression of bovine POU5F1

M J van Eijk, M A van Rooijen, S Modina, L Scesi, G Folkers, H T van Tol, M M Bevers, S R Fisher, H A Lewin, D Rakacolli, C Galli, C de Vaureix, A O Trounson, C L Mummery, F Gandolfi

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)1093-103
Number of pages11
JournalBiology of Reproduction
Volume60
Issue number5
Publication statusPublished - 1999

Keywords

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Biological Markers
  • Blotting, Northern
  • Cattle
  • Chromosome Mapping
  • Cloning, Molecular
  • DNA-Binding Proteins
  • Embryonic and Fetal Development
  • Female
  • Gene Expression Regulation, Developmental
  • Mice
  • Microscopy, Confocal
  • Molecular Sequence Data
  • Octamer Transcription Factor-3
  • Pregnancy
  • RNA, Messenger
  • Restriction Mapping
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stem Cells
  • Transcription Factors
  • Transfection

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