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Asymmetric cell divisions promote Notch-dependent epidermal differentiation

Nature volume 470, pages 353–358 (2011)Cite this article

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Abstract

Stem and progenitor cells use asymmetric cell divisions to balance proliferation and differentiation. Evidence from invertebrates shows that this process is regulated by proteins asymmetrically distributed at the cell cortex during mitosis: Par3–Par6–aPKC, which confer polarity, and Gαi–LGN/AGS3–NuMA–dynein/dynactin, which govern spindle positioning. Here we focus on developing mouse skin, where progenitor cells execute a switch from symmetric to predominantly asymmetric divisions concomitant with stratification. Using in vivo skin-specific lentiviral RNA interference, we investigate spindle orientation regulation and provide direct evidence that LGN (also called Gpsm2), NuMA and dynactin (Dctn1) are involved. In compromising asymmetric cell divisions, we uncover profound defects in stratification, differentiation and barrier formation, and implicate Notch signalling as an important effector. Our study demonstrates the efficacy of applying RNA interference in vivo to mammalian systems, and the ease of uncovering complex genetic interactions, here to gain insights into how changes in spindle orientation are coupled to establishing proper tissue architecture during skin development.

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Figure 1: Spindle orientation defects after LGN, Numa1 and Dctn1 depletion.
Figure 2: Impaired stratification in vitro and in vivo when asymmetric cell divisions are impaired.
Figure 3: Differentiation defects after LGN, Numa1 and Dctn1 depletion.
Figure 4: Loss of LGN or Numa1 impairs suprabasal Notch activation.
Figure 5: Genetic interaction between asymmetric cell division and Notch pathways.

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Acknowledgements

We thank N. Stokes, L. Polak and D. Oristian for their assistance in the mouse facility; S. Lanier and T. Gettys for providing AGS3 and Gαi antibodies; N. Gaiano and C. Cepko for constructs; D. Melton and T. Honjo for mice; S. Chai for Numb constructs; E. Ezhkova for sharing microarray data; and J. Knoblich for sharing unpublished results and reagents. We are grateful to M. Schober, D. Devenport, E. Ezratty, C. Luxenburg and members of the Fuchs laboratory for discussions and critical reading of the manuscript. We thank S. Mazel and the RU Flow Cytometry Resource Center for assistance with cell sorting, A. North and the RU Bioimaging Resource Center for assistance with image acquisition and the Comparative Biology Center (AAALAC accredited) for veterinary care of our mice. S.E.W. was supported by an American Cancer Society postdoctoral fellowship and S.B. was a Human Frontier Science Program postdoctoral fellow. 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. R01AR27883).

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  1. Howard Hughes Medical Institute, Laboratory of Mammalian Cell Biology and Development, The Rockefeller University, New York, 10065, New York, USA

    Scott E. Williams, Slobodan Beronja, H. Amalia Pasolli & Elaine Fuchs

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Contributions

S.E.W., E.F. and S.B. designed experiments. S.E.W. performed the experiments and analysed their raw data. H.A.P. conducted ultrastructural analyses. S.E.W. and S.B. performed lentiviral injections. S.E.W. and E.F. wrote the paper. All authors provided intellectual input, read and approved the manuscript.

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Correspondence to Elaine Fuchs.

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Williams, S., Beronja, S., Pasolli, H. et al. Asymmetric cell divisions promote Notch-dependent epidermal differentiation. Nature 470, 353–358 (2011). https://doi.org/10.1038/nature09793

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