Oct4A is a core component of the regulatory network of pluripotent cells, and by itself can reprogram neural stem cells into pluripotent cells in mice and humans. However, its role in defining totipotency and inducing pluripotency during embryonic development is still unclear. We genetically eliminated maternal Oct4A using a Cre/loxP approach in mouse and found that the establishment of totipotency was not affected, as shown by the generation of live pups. After complete inactivation of both maternal and zygotic Oct4A expression, the embryos still formed Oct4–GFP- and Nanog-expressing inner cell masses, albeit non-pluripotent, indicating that Oct4A is not a determinant for the pluripotent cell lineage separation. Interestingly, Oct4A-deficient oocytes were able to reprogram fibroblasts into pluripotent cells. Our results clearly demonstrate that, in contrast to its role in the maintenance of pluripotency, maternal Oct4A is not crucial for either the establishment of totipotency in embryos, or the induction of pluripotency in somatic cells using oocytes.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    & Totipotency, pluripotency and nuclear reprogramming. Adv. Biochem. Eng. Biotechnol. 114, 185–199 (2009).

  2. 2.

    , , , & A family of octamer-specific proteins present during mouse embryogenesis: evidence for germline-specific expression of an Oct factor. EMBO J. 8, 2543–2550 (1989).

  3. 3.

    & Oct-4: gatekeeper in the beginnings of mammalian development. Stem Cells 19, 271–278 (2001).

  4. 4.

    et al. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell 95, 379–391 (1998).

  5. 5.

    , , & Oct-3/4 and Sox2 regulate Oct-3/4 gene in embryonic stem cells. J. Biol. Chem. 280, 5307–5317 (2005).

  6. 6.

    , & Maternal control of early mouse development. Development 137, 859–870 (2010).

  7. 7.

    et al. Integration of external signaling pathways with the core transcriptional network in embryonic stem cells. Cell 133, 1106–1117 (2008).

  8. 8.

    et al. Transcriptional regulation of nanog by OCT4 and SOX2. J. Biol. Chem. 280, 24731–24737 (2005).

  9. 9.

    et al. Pluripotency governed by Sox2 via regulation of Oct3/4 expression in mouse embryonic stem cells. Nat. Cell Biol. 9, 625–635 (2007).

  10. 10.

    , , & Binding of Sp1 and Sp3 transcription factors to the Oct-4 gene promoter. Cell Mol. Biol. 45, 709–716 (1999).

  11. 11.

    & Stem cells, the molecular circuitry of pluripotency and nuclear reprogramming. Cell 132, 567–582 (2008).

  12. 12.

    et al. Oct4 is required for primordial germ cell survival. EMBO Rep. 5, 1078–1083 (2004).

  13. 13.

    et al. Oct4 expression is not required for mouse somatic stem cell self-renewal. Cell Stem Cell 1, 403–415 (2007).

  14. 14.

    , , , & The human OCT-4 isoformsdiffer in their ability to confer self-renewal. J. Biol. Chem. 281, 33554–33565 (2006).

  15. 15.

    et al. A novel variant of Oct3/4 gene in mouse embryonic stem cells. Stem Cell Res. 9, 69–76 (2012).

  16. 16.

    et al. OCT4B1, a novel spliced variant of OCT4, generates a stable truncated protein with a potential role in stress response. Cancer Lett. 309, 170–175 (2011).

  17. 17.

    et al. OCT4B1, a novel spliced variant of OCT4, is highly expressed in gastric cancer and acts as an antiapoptotic factor. Int. J. Cancer 128, 2645–2652 (2011).

  18. 18.

    et al. Maternal Oct-4 is a potential key regulator of the developmental competence of mouse oocytes. BMC Dev. Biol. 8, 97–110 (2008).

  19. 19.

    et al. A novel and critical role for Oct4 as a regulator of the maternal-embryonic transition. Plos One 3, e4109 (2008).

  20. 20.

    et al. Nanog safeguards pluripotency and mediates germline development. Nature 450, 1230–1234 (2007).

  21. 21.

    et al. Germline-specific expression of the Oct-4/green fluorescent protein (GFP) transgene in mice. Dev. Growth Differ. 41, 675–684 (1999).

  22. 22.

    , & Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat. Genet. 24, 372–376 (2000).

  23. 23.

    et al. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation. Cell 123, 917–929 (2005).

  24. 24.

    et al. Eset partners with Oct4 to restrict extraembryonic trophoblast lineage potential in embryonic stem cells. Genes Dev. 23, 2507–2520 (2009).

  25. 25.

    & Stochastic patterning in the mouse pre-implantation embryo. Development 134, 4219–4231 (2007).

  26. 26.

    et al. Ras-MAPK signaling promotes trophectoderm formation from embryonic stem cells and mouse embryos. Nat. Genet. 40, 921–926 (2008).

  27. 27.

    , , , & Maternal Cdx2 is dispensable for mouse development. Development 139, 3969–3972 (2012).

  28. 28.

    et al. Cdx2 is required for correct cell fate specification and differentiation of trophectoderm in the mouse blastocyst. Development 132, 2093–2102 (2005).

  29. 29.

    et al. Initiation of trophectoderm lineage specification in mouse embryos is independent of Cdx2. Development 137, 4159–4169 (2010).

  30. 30.

    et al. Gata3 regulates trophoblast development downstream of Tead4 and in parallel to Cdx2. Development 137, 395–403 (2010).

  31. 31.

    et al. Octamer and Sox elements are required for transcriptional cis regulation of Nanog gene expression. Mol. Cell Biol. 25, 2475–2485 (2005).

  32. 32.

    , , & Allele-specific expression of imprinted genes in mouse migratory primordial germ cells. Mech. Dev. 115, 157–160 (2002).

  33. 33.

    & Directing reprogramming to pluripotency by transcription factors. Curr. Opin. Genet. Dev. 22, 1–7 (2012).

  34. 34.

    et al. The nuclear receptor Nr5a2 can replace Oct4 in the reprogramming of murine somatic cells to pluripotent cells. Cell Stem Cell 6, 167–174 (2010).

  35. 35.

    & Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676 (2006).

  36. 36.

    et al. Orphan nuclear receptor LRH-1 is required to maintain Oct4 expression at the epiblast stage of embryonic development. Mol. Cell Biol. 25, 3492–3505 (2005).

  37. 37.

    & A genome-wide screen in EpiSCs identifies Nr5a nuclear receptors as potent inducers of ground state pluripotency. Development 137, 3185–3192 (2010).

  38. 38.

    et al. Expression of the T-box family genes, Tbx1-Tbx5, during early mouse development. Dev. Dyn. 206, 379–390 (1996).

  39. 39.

    et al. Role of TBX1 in human del22q11.2 syndrome. Lancet 362, 1366–1373 (2003).

  40. 40.

    , , & Sheep cloned by nuclear transfer from a cultured cell line. Nature 380, 64–66 (1996).

  41. 41.

    et al. Expression of Cre recombinase in mouse oocytes: a means to study maternal effect genes. Genesis 26, 110–112 (2000).

  42. 42.

    , & Differential oocyte-specific expression of Cre recombinase activity in GDF-9-iCre, Zp3cre, and Msx2Cre transgenic mice. Biol. Reprod. 71, 1469–1474 (2004).

  43. 43.

    , & Single cell detection of beta-thalassaemia mutations using silver stained SSCP analysis: an application for preimplantation diagnosis. Mol. Hum. Reprod. 3, 693–698 (1997).

  44. 44.

    Manipulating The Mouse Embryo : A Laboratory Manual 2nd edn (Cold Spring Harbor Laboratory Press, 1994).

  45. 45.

    , , & Oct-4 transcription factor is differentially expressed in the mouse embryo during establishment of the first two extraembryonic cell lineages involved in implantation. Dev. Biol. 166, 259–267 (1994).

  46. 46.

    et al. Variable reprogramming of the pluripotent stem cell marker Oct4 in mouse clones: distinct developmental potentials in different culture environments. Stem Cells 23, 1089–1104 (2005).

  47. 47.

    , , & Preimplantation development of mouse embryos in KSOM: augmentation by amino acids and analysis of gene expression. Mol. Reprod. Dev. 41, 232–238 (1995).

  48. 48.

    Manipulating The Mouse Embryo : A Laboratory Manual 3rd edn (Cold Spring Harbor Laboratory Press, 2003).

  49. 49.

    , & Genetic and spectrally distinct in vivo imaging: embryonic stem cells and mice with widespread expression of a monomeric red fluorescent protein. BMC Biotechnol. 5, 20–30 (2005).

  50. 50.

    et al. Mitochondrial gene replacement in primate offspring and embryonic stem cells. Nature 461, 367–372 (2009).

  51. 51.

    , & Derivation of mouse embryonic stem cells. Nat. Protoc. 1, 2082–2087 (2006).

  52. 52.

    et al. Pluripotent stem cells induced from adult neural stem cells by reprogramming with two factors. Nature 454, 646–650 (2008).

  53. 53.

    et al. Efficient derivation of pluripotent stem cells from siRNA-mediated Cdx2-deficient mouse embryos. Stem Cells Dev. 20, 485–493 (2011).

  54. 54.

    et al. Generation of healthy mice from gene-corrected disease-specific induced pluripotent stem cells. PLoS Biol. 9, e1001099 (2011).

Download references


We thank J. Mueller-Keuker, M. Preusser and N. Stengel for assistance in preparing the manuscript and A. Malapetsas for proofreading the manuscript. We thank K. Huebner for her technical help on immunocytochemistry and B. Scháfer for her assistance on histology work. The authors of this manuscript bear sole responsibility for the content presented, which does not necessarily represent the official views of the Eunice Kennedy Shriver National Institute of Child Health & Human Development or the National Institutes of Health. This research was supported by the Max Planck Society, DFG grants DFG SI 1695/1-2 (SPP1356) and SCHO 340/7-1, and grant NIH R01HD059946-01 from the Eunice Kennedy Shriver National Institute of Child Health & Human Development.

Author information

Author notes

    • Vittorio Sebastiano
    •  & Nishant Singhal

    Present addresses: Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, 1050 Arastradero Road, Palo Alto, California 94304, USA (V.S.); Department of Neurosciences, University of California San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA (N.S.)


  1. Max Planck Institute for Molecular Biomedicine, Department of Cell and Developmental Biology, Röntgenstrasse 20, 48149 Münster, Germany

    • Guangming Wu
    • , Dong Han
    • , Yu Gong
    • , Vittorio Sebastiano
    • , Luca Gentile
    • , Nishant Singhal
    • , Kenjiro Adachi
    • , Gerrit Fischedick
    • , Claudia Ortmeier
    • , Martina Sinn
    • , Martina Radstaak
    •  & Hans R. Schöler
  2. Russian Academy of Science, Institute of Cytology, 4 Tikhoretski Avenue, 194064, St Petersburg, Russia

    • Alexey Tomilin
  3. University of Münster, Medical Faculty, Domagkstr. 3, 48149 Münster, Germany

    • Hans R. Schöler


  1. Search for Guangming Wu in:

  2. Search for Dong Han in:

  3. Search for Yu Gong in:

  4. Search for Vittorio Sebastiano in:

  5. Search for Luca Gentile in:

  6. Search for Nishant Singhal in:

  7. Search for Kenjiro Adachi in:

  8. Search for Gerrit Fischedick in:

  9. Search for Claudia Ortmeier in:

  10. Search for Martina Sinn in:

  11. Search for Martina Radstaak in:

  12. Search for Alexey Tomilin in:

  13. Search for Hans R. Schöler in:


G.W. designed and executed experiments as well as writing the manuscript. D.H., Y.G., V.S., L.G., N.S., K.A., G.F., C.O., M.S., M.R. and A.T. executed experiments, collected data and prepared reagents. H.R.S. provided the study concept and funding, and edited the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Hans R. Schöler.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Information

Excel files

  1. 1.

    Supplementary Tables 1–4

    Supplementary Information

  2. 2.

    Supplementary Table 5

    Supplementary Information


  1. 1.

    Time-lapse recording of in vitro development of Oct4A-null 8-cell embryo.

    A biopsied and genotyped morula with maternal and zygotic Oct4A-null was cultured on MEFs in ESC medium and observed on the stage of a microscope with an incubation chamber (TOKAI HIT, Japan) filled with 5% CO2 in air and maintained at 37 °C. Brightfield pictures were taken every 5 min for 4 days and were compiled into a movie with 24 frames per second. The video demonstrates that Oct4A-null embryos initiated cavitation and formed grossly normal-looking blastocysts with distinct ICM. However, immunostaining of the outgrowth (Fig. 3a) showed cytoplasmic localization of Nanog as well as fragmentation of nuclei.

  2. 2.

    Time-lapse confocal recording revealed activation of Oct4-GFP expression in maternal-knockout and maternal/zygotic-knockout embryos.

    Twelve 2-cell embryos from the mating of Oct4flox/flox/ZP3Cre/+ female mice with Oct4A+/Δ/Oct4−GFP+/+ male mice and 4 embryos (#1, 3, 4 and 8) from the mating of Oct4flox/flox female mice with Oct4A+/Δ/Oct4GFP+/+ male mice were placed in KSOMAA in a glass bottom dish with the same condition as Supplementary Video 1 for confocal examination with 488 nm laser. A confocal picture had been taken every 10 min for 3 days and was compiled into a movie with 24 frames per second. The video demonstrated that regardless of the genotype, all embryos activated Oct4-GFP at around E2.5in a timely fashion, as did wild-type embryos. The genotype of each embryo is shown in Fig. S3a.

  3. 3.

    Brightfield time-lapse recording of the same embryos at the same time point as Supplementary Video 2.

    This video was used to monitor the developmental stage of the embryos and to trace the position of individual embryos for genotype determination.

About this article

Publication history






Further reading