Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells

Abstract

Cell fate during development is defined by transcription factors that act as molecular switches to activate or repress specific gene expression programmes. The POU transcription factor Oct-3/4 (encoded by Pou5f1) is a candidate regulator in pluripotent and germline cells1,2,3,4 and is essential for the initial formation of a pluripotent founder cell population in the mammalian embryo5. Here we use conditional expression and repression in embryonic stem (ES) cells to determine requirements for Oct-3/4 in the maintenance of developmental potency. Although transcriptional determination has usually been considered as a binary on-off control system, we found that the precise level of Oct-3/4 governs three distinct fates of ES cells. A less than twofold increase in expression causes differentiation into primitive endoderm and mesoderm. In contrast, repression of Oct-3/4 induces loss of pluripotency and dedifferentiation to trophectoderm. Thus a critical amount of Oct-3/4 is required to sustain stem-cell self-renewal, and up- or downregulation induce divergent developmental programmes. Our findings establish a role for Oct-3/4 as a master regulator of pluripotency that controls lineage commitment and illustrate the sophistication of critical transcriptional regulators and the consequent importance of quantitative analyses.

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Figure 1: Generation of ES cells with variable Oct-3/4 expression.
Figure 2: Differentiation of ZHTc6 and ZHBTc4 ES cells induced by modulation of Oct-3/4 expression.
Figure 3: Morphological differentiation of ZHBTc4 ES cells on repression of Oct-3/4.
Figure 4: Comparison of gene-expression pattern during differentiation of ZHBTc4 and ZHTc6 ES cells.
Figure 5: TS cell derivation from ZHBTc4 ES cells.
Figure 6: Functions of Oct-3/4 in pluripotent stem cells.

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Acknowledgements

We thank S. Nishikawa for access to time-lapse recording facilities and H. Schöler and H. Hamada for anti-Oct-3/4 antisera. We acknowledge F. Stehnouse for technical assistance. This work was supported by the Biotechnology and Biological Sciences Research Council of the United Kingdom, The Ministry of Education, Science and Culture of Japan, and a Uehara Memorial Foundation Award to H.N.

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Correspondence to Hitoshi Niwa or Austin G. Smith.

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Niwa, H., Miyazaki, Ji. & Smith, A. Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat Genet 24, 372–376 (2000). https://doi.org/10.1038/74199

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