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MCPH1 regulates the neuroprogenitor division mode by coupling the centrosomal cycle with mitotic entry through the Chk1–Cdc25 pathway

Nature Cell Biology volume 13, pages 1325–1334 (2011)Cite this article

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Abstract

Primary microcephaly 1 is a neurodevelopmental disorder caused by mutations in the MCPH1 gene, whose product MCPH1 (also known as microcephalin and BRIT1) regulates DNA-damage response. Here we show that Mcph1 disruption in mice results in primary microcephaly, mimicking human MCPH1 symptoms, owing to a premature switching of neuroprogenitors from symmetric to asymmetric division. MCPH1-deficiency abrogates the localization of Chk1 to centrosomes, causing premature Cdk1 activation and early mitotic entry, which uncouples mitosis and the centrosome cycle. This misorients the mitotic spindle alignment and shifts the division plane of neuroprogenitors, to bias neurogenic cell fate. Silencing Cdc25b, a centrosome substrate of Chk1, corrects MCPH1-deficiency-induced spindle misalignment and rescues the premature neurogenic production in Mcph1-knockout neocortex. Thus, MCPH1, through its function in the Chk1–Cdc25–Cdk1 pathway to couple the centrosome cycle with mitosis, is required for precise mitotic spindle orientation and thereby regulates the progenitor division mode to maintain brain size.

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Figure 1: The microcephaly of Mcph1-del mice.
Figure 2: Neocortical developmental analysis of Mcph1-del mice.
Figure 3: Characterization of Mcph1-knockout neural progenitors in vitro and in vivo.
Figure 4: In vivo analyses of the cell division mode of neuroprogenitors.
Figure 5: Analysis of cell-cycle-dependent Chk1 centrosome localization, premature mitotic entry and centrosome maturation in primary neuroprogenitors.
Figure 6: Analysis of premature mitotic entry and centrosome maturation in primary neuroprogenitors.
Figure 7: Characterization of mitotic spindle defects in Mcph1-del neuroprogenitors and MEFs by Chk1 and/or Cdc25b knockdown.
Figure 8: In vivo silencing of Chk1 and Cdc25b in the neocortex by in utero electroporation.

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Acknowledgements

We thank L. Frappart for the advice and the discussion on histological analysis and Z. Xu for the introduction into the in utero electroporation technology. We are grateful to M. Baldauf for his histological assistance, M. Welzel for her technical support and D. Galendo, C. Birch-Hirschfeld and C. Mueller for their assistance in the maintenance of the animal colonies. We are also grateful to M. Trajkovic-Arsic for the advice on the in situ hybridization techniques and A. Gompf for the FACS. We also would like to thank P. Herrlich, H. Heuer, T. Li and Y. Yang for their critical reading of the manuscript. Further thanks go to E. Stoeckl for editing the manuscript. We are also grateful to many other members of our laboratory for the helpful discussions. R.G., M.S. and Z.W.Z. are supported by a fellowship of the Leibniz Graduate School on Ageing (LGSA) Programme. P-O.F. is supported by an Emmy Noether Grant from the Deutschen Forschungsgemeinschaft (DFG). Z-Q.W. is supported in part by the DFG, Germany.

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  1. Ralph Gruber

    Present address: Present address: Cancer Research UK—London Research Institute, 44 Lincoln’s Inn Fields, London WC2A 3LY, UK,

  2. Zhongwei Zhou and Mikhail Sukchev: These authors contributed equally to this work

Authors and Affiliations

  1. Leibniz Institute for Age Research—Fritz Lipmann Institute (FLI), Beurtenbergstrasse 11, 07745 Jena, Germany

    Ralph Gruber, Zhongwei Zhou, Mikhail Sukchev, Tjard Joerss & Zhao-Qi Wang

  2. Institute of Molecular Cell Biology, Centre for Molecular Biomedicine (CMB), Hans-Knöll-Strasse 2, 07745 Jena, Germany

    Pierre-Olivier Frappart

  3. Faculty of Biology and Pharmacy, Friedrich Schiller University of Jena, Beurtenbergstrasse 11, 07745 Jena, Germany

    Zhao-Qi Wang

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Contributions

R.G. carried out most of the experiments, analysed data and prepared the figures and the manuscript; Z.W.Z. carried out in situ hybridization, in utero electroporation, immunoblot analysis and analysed data; M.S. carried out gene targeting in embryonic stem cells and analysed gene expression; P-O.F. assisted with neurosphere experiments; T.J. contributed to mouse colony maintenance; Z-Q.W. designed experiments, analysed data and composed the manuscript.

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Correspondence to Ralph Gruber or Zhao-Qi Wang.

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Gruber, R., Zhou, Z., Sukchev, M. et al. MCPH1 regulates the neuroprogenitor division mode by coupling the centrosomal cycle with mitotic entry through the Chk1–Cdc25 pathway. Nat Cell Biol 13, 1325–1334 (2011). https://doi.org/10.1038/ncb2342

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