title: Effect of ploidy and parental genome composition on expression of Oct-4 protein in mouse embryos.
abstract: The transcription factor Oct-4 is expressed in germ cells and also is considered as a marker for pluripotency of stem cells. We first examined dynamics of Oct-4 protein expression during preimplantation development using both Western blot analysis, and immunofluorescence staining. We show that intact Oct-4 protein is not detected in either ovulated mature oocytes, or in zygotes and 2-4-cell embryos, which are the only known totipotent cell types in mammals. This finding is unexpected, since Oct-4 has been proposed to play a role in the control of totipotency. The results suggest that Oct-4 is not indispensable for fertilization and early cleavage. Rather, expression of Oct-4 protein is first detected in the nuclei of 8-16 cell morula, increases in early blastocysts, and declines in late blastocysts, in which most Oct-4 protein is confined to the inner cell mass (ICM) region, consistent with previous findings. We further compared Oct-4 protein expression in diploid and tetraploid blastocysts derived from normal fertilization or parthenogenesis, as well as expression in diploid androgenetic blastocysts. Expression levels and localization of Oct-4 protein are similar in both diploid and tetraploid early blastocysts, regardless of whether blastocysts are derived from fertilization or parthenogenesis. Androgenetic diploid blastocysts also express similar levels of Oct-4. Late blastocysts generated by both fertilization and parthenogenesis show a similar pattern of Oct-4 expression, suggesting that paternal genome activation is not required for Oct-4 expression. Expression of Oct-4 protein does not differ between diploid and tetraploid embryos, indicating that tetraploidy does not influence Oct-4 expression. Thus, expression of Oct-4 protein is initiated at morula stage in preimplantation embryos and completely controlled by a mechanism activated in oocytes. Downregulation of Oct-4 expression coincides with differentiation of trophectoderm. Similar profiles of Oct-4 expression observed in embryos with different ploidy and genome composition, are suggestive of Oct-4 being necessary but not sufficient for developmental potency.
authors.affiliation: Department of Obstetrics/Gynecology, Women and Infants Hospital, Brown Medical School, Providence, RI 02905, USA. lliu@mbl.edu
authors.forename: Lin
authors.initials: L
authors.lastname: Liu
authors.forename: Eva
authors.initials: E
authors.lastname: Czerwiec
authors.forename: David L
authors.initials: DL
authors.lastname: Keefe
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D001757:Blastomeres / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology*; D051379:Mice; D009028:Morula / Q000378:metabolism*; D050814:Octamer Transcription Factor-3; D010312:Parthenogenesis / Q000502:physiology; D011123:Polyploidy; D011247:Pregnancy; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D014327:Trophoblasts / Q000378:metabolism; D015053:Zygote / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D014157:Transcription Factors