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Dynamic maintenance of asymmetric meiotic spindle position through Arp2/3-complex-driven cytoplasmic streaming in mouse oocytes

Nature Cell Biology volume 13, pages 1252–1258 (2011)Cite this article

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Abstract

Mature mammalian oocytes are poised for completing meiosis II (MII) on fertilization by positioning the spindle close to an actomyosin-rich cortical cap1,2,3. Here, we show that the Arp2/3 complex localizes to the cortical cap in a Ran-GTPase-dependent manner and nucleates actin filaments in the cortical cap and a cytoplasmic actin network. Inhibition of Arp2/3 activity leads to rapid dissociation of the spindle from the cortex. Live-cell imaging and spatiotemporal image correlation spectroscopy analysis reveal that actin filaments flow continuously away from the Arp2/3-rich cortex, driving a cytoplasmic streaming expected to exert a net pushing force on the spindle towards the cortex. Arp2/3 inhibition not only diminishes this actin flow and cytoplasmic streaming but also enables a reverse streaming driven by myosin-II-based cortical contraction, moving the spindle away from the cortex. Thus, the asymmetric MII spindle position is dynamically maintained as a result of balanced forces governed by the Arp2/3 complex.

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Figure 1: Inhibition of Arp2/3-complex activity disrupts asymmetric MII spindle position.
Figure 2: Ran signalling regulates cortical localization of the Arp2/3 complex.
Figure 3: The Arp2/3 complex is required for most F-actin assembly in the cortical cap and for myosin-II ring maintenance.
Figure 4: Cytoplasmic streaming powered by Arp2/3-complex-dependent actin flow generates a net pushing force on the spindle.
Figure 5: Myosin-II-dependent cortical cap contraction drives the MII spindle away from the cortex in the absence of Arp2/3 activity.

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Acknowledgements

We thank W. M. Bement (University of Wisconsin, USA) for providing pCS2+–UtrCH–GFP plasmid; J. Bamburg (Colorado State University, USA) for providing anti-cofilin and phos-cofilin antibodies; H. Cartwright (Stowers Institute, USA) for microfabricated wells for oocyte imaging; and M. Durnin and K. Westfahl (both Stowers Institute, USA) for technical assistance and mice maintenance. This work was supported in part by NIH grant P01 GM 066311.

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Authors and Affiliations

  1. Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, Missouri 64110, USA

    Kexi Yi, Jay R. Unruh, Brian D. Slaughter, Boris Rubinstein & Rong Li

  2. Department of Obstetrics and Gynecology and Reproductive Biology, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA

    Manqi Deng

  3. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas 66160, 3901 Rainbow Boulevard, USA

    Rong Li

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Contributions

K.Y. and R.L. designed the experiments, interpreted results and prepared the manuscript; K.Y. carried out all of the experiments; J.R.U. carried out STICS analysis with assistance from B.D.S. and also contributed to other image analysis; M.D. assisted in the initial experimental set-up. B.R. carried out the numerical simulations; R.L. conceived and supervised the project.

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Correspondence to Rong Li.

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The authors declare no competing financial interests.

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Yi, K., Unruh, J., Deng, M. et al. Dynamic maintenance of asymmetric meiotic spindle position through Arp2/3-complex-driven cytoplasmic streaming in mouse oocytes. Nat Cell Biol 13, 1252–1258 (2011). https://doi.org/10.1038/ncb2320

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