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Par3–mInsc and Gαi3 cooperate to promote oriented epidermal cell divisions through LGN

Nature Cell Biology volume 16, pages 758769 (2014) | Download Citation

Abstract

Asymmetric cell divisions allow stem cells to balance proliferation and differentiation. During embryogenesis, murine epidermis expands rapidly from a single layer of unspecified basal layer progenitors to a stratified, differentiated epithelium. Morphogenesis involves perpendicular (asymmetric) divisions and the spindle orientation protein LGN, but little is known about how the apical localization of LGN is regulated. Here, we combine conventional genetics and lentiviral-mediated in vivo RNAi to explore the functions of the LGN-interacting proteins Par3, mInsc and Gαi3. Whereas loss of each gene alone leads to randomized division angles, combined loss of Gnai3 and mInsc causes a phenotype of mostly planar divisions, akin to loss of LGN. These findings lend experimental support for the hitherto untested model that Par3–mInsc and Gαi3 act cooperatively to polarize LGN and promote perpendicular divisions. Finally, we uncover a developmental switch between delamination-driven early stratification and spindle-orientation-dependent differentiation that occurs around E15, revealing a two-step mechanism underlying epidermal maturation.

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Acknowledgements

We thank N. Stokes, D. Oristian and A. Aldeguer (Fuchs laboratory) and T. Anthony Curtis (Williams laboratory), for their expert technical assistance. We thank K. Byrd, K. Lough, and members of the Williams and Fuchs laboratories for critical reading of the manuscript and K. Lough for valuable input into the model presented in Fig. 8. We are grateful to S. Ohno and T. Hirose (both at Yokohama City University of Medicine, Japan) for sharing the Pard3 floxed mouse line. S.E.W. was supported by an American Cancer Society postdoctoral fellowship and E.F. is an investigator in the Howard Hughes Medical Institute. Work in the Fuchs laboratory was supported by a grant from the National Institutes of Health (E.F. R37-27883).

Author information

Author notes

    • Maria Pia Postiglione

    Present address: Institute of Science and Technology Austria, Am Campus 1, A-3400 Klosterneuburg, Austria.

Affiliations

  1. Howard Hughes Medical Institute, Laboratory of Mammalian Cell Biology & Development, The Rockefeller University, 1230 York Avenue, Box 300, New York, New York 10065, USA

    • Scott E. Williams
    •  & Elaine Fuchs
  2. Department of Pathology & Laboratory Medicine, Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina 27599, USA

    • Scott E. Williams
    •  & Lyndsay A. Ratliff
  3. Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohrgasse 3, 1030 Vienna, Austria

    • Maria Pia Postiglione
    •  & Juergen A. Knoblich

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Contributions

S.E.W. designed and conducted experiments and analysed the data under the supervision of E.F. L.A.R. performed the imaging and analysis for the lineage tracing experiments. M.P.P. and J.A.K. provided mInsc mice before publication. S.E.W. and E.F. wrote the manuscript. All authors critically read and contributed to the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Elaine Fuchs.

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DOI

https://doi.org/10.1038/ncb3001

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