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Developmental roles for Srf, cortical cytoskeleton and cell shape in epidermal spindle orientation

Nature Cell Biology volume 13, pages 203–214 (2011)Cite this article

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Abstract

During development, a polarized epidermal sheet undergoes stratification and differentiation to produce the skin barrier. Through mechanisms that are poorly understood, the process involves actin dynamics, spindle reorientation and Notch signalling. To elucidate how epidermal embryogenesis is governed, we conditionally targeted serum response factor (Srf), a transcription factor that is essential for epidermal differentiation. Unexpectedly, previously ascribed causative defects are not responsible for profoundly perturbed embryonic epidermis. Seeking the mechanism for this, we identified actins and their regulators that were downregulated after ablation. Without Srf, cells exhibit a diminished cortical network and in mitosis, they fail to round up, features we recapitulate with low-dose actin inhibitors in vivo and shRNA-knockdown in vitro. Altered concomitantly are phosphorylated ERM and cortical myosin-IIA, shown in vitro to establish a rigid cortical actomyosin network and elicit critical shape changes. We provide a link between these features and Srf loss, and we show that the process is physiologically relevant in skin, as reflected by defects in spindle orientation, asymmetric cell divisions, stratification and differentiation.

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Figure 1: Srf is expressed in embryonic basal cells and when absent, morphological abnormalities originate at the basal to suprabasal juncture.
Figure 2: Alterations in biochemical markers of the basal–suprabasal switch are obvious by E17.5 in Srf-null epidermis.
Figure 3: Cell adhesion remains intact during embryonic development in Srf-cKO skin.
Figure 4: Conditional Srf ablation results in an early reduction in cortical actin, which seems to be directly responsible for an associated ability of early mitotic cells to round up during mitosis.
Figure 5: Srf-cKO early mitotic cells are defective in enriching and activating ERM proteins at the actin cortex and in recruiting myosin-IIA.
Figure 6: Actin and actin regulators downregulated in Srf-deficient basal cells are required for keratinocytes to undergo proper cortical and cell shape changes during mitosis.
Figure 7: Mitotic abnormalities in spindle orientation and LGN–NuMA localization in E16.5 Srf-cKO basal cells.

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Acknowledgements

We thank the RU Facilities for Bioimaging (A. North) and FACS (S. Mazel) staff for their technical support; we thank N. Stokes for her assistance in the mouse facility and the Comparative Biology Center (CBC) staff for their help in veterinary care and health of our mice. We are grateful to E. Ezratty, S. Beronja, A. R. Folgueras, M. Nikolova and other members of the Fuchs laboratory for helpful discussions, critical reading of the manuscript and experimental assistance. C.L. is a Tri-Institutional Starr Stem Cell Scholars Posdoctoral Fellow. S.E.W. is an American Cancer Society Postdoctoral Fellow. 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, New York 10065, USA

    Chen Luxenburg, H. Amalia Pasolli, Scott E. Williams & Elaine Fuchs

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  1. Chen Luxenburg
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C.L. and H.A.P. carried out the experiments and analysed the raw data. S.E.W. contributed reagents and experimental suggestions. C.L. and E.F. wrote the manuscript. E.F. supervised the project.

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Correspondence to Elaine Fuchs.

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Luxenburg, C., Amalia Pasolli, H., Williams, S. et al. Developmental roles for Srf, cortical cytoskeleton and cell shape in epidermal spindle orientation. Nat Cell Biol 13, 203–214 (2011). https://doi.org/10.1038/ncb2163

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