A contractile nuclear actin network drives chromosome congression in oocytes

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Abstract

Chromosome capture by microtubules is widely accepted as the universal mechanism of spindle assembly in dividing cells. However, the observed length of spindle microtubules and computer simulations of spindle assembly predict that chromosome capture is efficient in small cells, but may fail in cells with large nuclear volumes such as animal oocytes. Here we investigate chromosome congression during the first meiotic division in starfish oocytes. We show that microtubules are not sufficient for capturing chromosomes. Instead, chromosome congression requires actin polymerization. After nuclear envelope breakdown, we observe the formation of a filamentous actin mesh in the nuclear region, and find that contraction of this network delivers chromosomes to the microtubule spindle. We show that this mechanism is essential for preventing chromosome loss and aneuploidy of the egg—a leading cause of pregnancy loss and birth defects in humans.

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Figure 1: Chromosomes congress without attachment to meiotic asters.
Figure 2: Chromosome congression is independent of microtubules but relies on actin polymerization.
Figure 3: Actin polymerization initiates at NEBD.
Figure 4: Actin polymerizes into a contractile network in the nuclear region that moves chromosomes to the animal pole.
Figure 5: F-actin-stabilizing drugs prevent chromosome movement and contraction of the actin network.

References

  1. 1

    Gadde, S. & Heald, R. Mechanisms and molecules of the mitotic spindle. Curr. Biol. 14, R797–R805 (2004)

  2. 2

    Rusan, N. M., Fagerstrom, C. J., Yvon, A. M. & Wadsworth, P. Cell cycle-dependent changes in microtubule dynamics in living cells expressing green fluorescent protein-α tubulin. Mol. Biol. Cell 12, 971–980 (2001)

  3. 3

    Zhai, Y., Kronebusch, P. J., Simon, P. M. & Borisy, G. G. Microtubule dynamics at the G2/M transition: abrupt breakdown of cytoplasmic microtubules at nuclear envelope breakdown and implications for spindle morphogenesis. J. Cell Biol. 135, 201–214 (1996)

  4. 4

    Kirschner, M. & Mitchison, T. Beyond self-assembly: from microtubules to morphogenesis. Cell 45, 329–342 (1986)

  5. 5

    Kline-Smith, S. L. & Walczak, C. E. Mitotic spindle assembly and chromosome segregation: refocusing on microtubule dynamics. Mol. Cell 15, 317–327 (2004)

  6. 6

    Holy, T. E. & Leibler, S. Dynamic instability of microtubules as an efficient way to search in space. Proc. Natl Acad. Sci. USA 91, 5682–5685 (1994)

  7. 7

    Belmont, L. D., Hyman, A. A., Sawin, K. E. & Mitchison, T. J. Real-time visualization of cell cycle-dependent changes in microtubule dynamics in cytoplasmic extracts. Cell 62, 579–589 (1990)

  8. 8

    Piehl, M. & Cassimeris, L. Organization and dynamics of growing microtubule plus ends during early mitosis. Mol. Biol. Cell 14, 916–925 (2003)

  9. 9

    Rieder, C. L. & Alexander, S. P. Kinetochores are transported poleward along a single astral microtubule during chromosome attachment to the spindle in newt lung cells. J. Cell Biol. 110, 81–95 (1990)

  10. 10

    Carazo-Salas, R. E. & Karsenti, E. Long-range communication between chromatin and microtubules in Xenopus egg extracts. Curr. Biol. 13, 1728–1733 (2003)

  11. 11

    Tulu, U. S., Rusan, N. M. & Wadsworth, P. Peripheral, non-centrosome-associated microtubules contribute to spindle formation in centrosome-containing cells. Curr. Biol. 13, 1894–1899 (2003)

  12. 12

    Wollman, R. et al. Efficient chromosome capture requires a bias in the ‘search-and-capture’ process during mitotic-spindle assembly. Curr. Biol. 15, 828–832 (2005)

  13. 13

    Zhang, Q. Y., Tamura, M., Uetake, Y., Washitani-Nemoto, S. & Nemoto, S. Regulation of the paternal inheritance of centrosomes in starfish zygotes. Dev. Biol. 266, 190–200 (2004)

  14. 14

    Lenart, P. et al. Nuclear envelope breakdown in starfish oocytes proceeds by partial NPC disassembly followed by a rapidly spreading fenestration of nuclear membranes. J. Cell Biol. 160, 1055–1068 (2003)

  15. 15

    Miyazaki, A., Kamitsubo, E. & Nemoto, S. I. Premeiotic aster as a device to anchor the germinal vesicle to the cell surface of the presumptive animal pole in starfish oocytes. Dev. Biol. 218, 161–171 (2000)

  16. 16

    Gerlich, D. et al. Global chromosome positions are transmitted through mitosis in mammalian cells. Cell 112, 751–764 (2003)

  17. 17

    Stricker, S. A. & Schatten, G. The cytoskeleton and nuclear disassembly during germinal vesicle breakdown in starfish oocytes. Dev. Growth Differ. 33, 163–171 (1991)

  18. 18

    Terasaki, M. Redistribution of cytoplasmic components during germinal vesicle breakdown in starfish oocytes. J. Cell Sci. 107, 1797–1805 (1994)

  19. 19

    Pang, K. M., Lee, E. & Knecht, D. A. Use of a fusion protein between GFP and an actin-binding domain to visualize transient filamentous-actin structures. Curr. Biol. 8, 405–408 (1998)

  20. 20

    Heil-Chapdelaine, R. A. & Otto, J. J. Characterization of changes in F-actin during maturation of starfish oocytes. Dev. Biol. 177, 204–216 (1996)

  21. 21

    Visegradi, B., Lorinczy, D., Hild, G., Somogyi, B. & Nyitrai, M. The effect of phalloidin and jasplakinolide on the flexibility and thermal stability of actin filaments. FEBS Lett. 565, 163–166 (2004)

  22. 22

    Weber, K. L., Sokac, A. M., Berg, J. S., Cheney, R. E. & Bement, W. M. A microtubule-binding myosin required for nuclear anchoring and spindle assembly. Nature 431, 325–329 (2004)

  23. 23

    Waterman-Storer, C. et al. Microtubules remodel actomyosin networks in Xenopus egg extracts via two mechanisms of F-actin transport. J. Cell Biol. 150, 361–376 (2000)

  24. 24

    Szent-Gyorgyi, A. Chemistry of Muscular Contraction (Academic, New York, 1951)

  25. 25

    Spicer, S. S. The clearing response of actomyosin to adenosinetriphosphate. J. Biol. Chem. 199, 289–300 (1952)

  26. 26

    Gard, D. L. Microtubule organization during maturation of Xenopus oocytes: assembly and rotation of the meiotic spindles. Dev. Biol. 151, 516–530 (1992)

  27. 27

    Ryabova, L. V., Betina, M. I. & Vassetzky, S. G. Influence of cytochalasin B on oocyte maturation in Xenopus laevis. Cell Differ. 19, 89–96 (1986)

  28. 28

    Gard, D. L., Cha, B. J. & Roeder, A. D. F-actin is required for spindle anchoring and rotation in Xenopus oocytes: a re-examination of the effects of cytochalasin B on oocyte maturation. Zygote 3, 17–26 (1995)

  29. 29

    Longo, F. J. & Chen, D. Y. Development of cortical polarity in mouse eggs: involvement of the meiotic apparatus. Dev. Biol. 107, 382–394 (1985)

  30. 30

    Leader, B. et al. Formin-2, polyploidy, hypofertility and positioning of the meiotic spindle in mouse oocytes. Nature Cell Biol. 4, 921–928 (2002)

  31. 31

    Maro, B. & Verlhac, M. H. Polar body formation: new rules for asymmetric divisions. Nature Cell Biol. 4, E281–E283 (2002)

  32. 32

    Peter, M. et al. The APC is dispensable for first meiotic anaphase in Xenopus oocytes. Nature Cell Biol. 3, 83–87 (2001)

  33. 33

    Wassmann, K., Niault, T. & Maro, B. Metaphase I arrest upon activation of the Mad2-dependent spindle checkpoint in mouse oocytes. Curr. Biol. 13, 1596–1608 (2003)

  34. 34

    Hassold, T. & Hunt, P. To err (meiotically) is human: the genesis of human aneuploidy. Nature Rev. Genet. 2, 280–291 (2001)

  35. 35

    Faire, K. et al. E-MAP-115 (ensconsin) associates dynamically with microtubules in vivo and is not a physiological modulator of microtubule dynamics. J. Cell Sci. 112, 4243–4255 (1999)

  36. 36

    Hinkle, B. et al. Chromosomal association of Ran during meiotic and mitotic divisions. J. Cell Sci. 115, 4685–4693 (2002)

  37. 37

    Heald, R. et al. Self-organization of microtubules into bipolar spindles around artificial chromosomes in Xenopus egg extracts. Nature 382, 420–425 (1996)

  38. 38

    Strickland, L. et al. Light microscopy of echinoderm embryos. Methods Cell Biol. 74, 371–409 (2004)

  39. 39

    Bacher, C. P., Reichenzeller, M., Athale, C., Herrmann, H. & Eils, R. 4-D single particle tracking of synthetic and proteinaceous microspheres reveals preferential movement of nuclear particles along chromatin-poor tracks. BMC Cell Biol. 5, 45 (2004)

  40. 40

    Hand, A. R. in Introduction to Biophysical Methods for Protein and Nucleic Acid Research (eds Glasel, J. & Deutscher, M.) 205–260 (Academic, New York, 1995)

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Acknowledgements

Part of this work was performed at the Marine Biological Laboratory (MBL) in Woods Hole, supported by summer research fellowships from Nikon Inc. and the E. and M. Spiegel, F.B. and B.G. Bang, L.B. Lehmann, R.D. Allen and H.W. Rand foundations to J.E. Martin Hoppe and Leica Microsystems in Mannheim are gratefully acknowledged for providing equipment at the MBL. P.L. was supported by a predoctoral fellowship from the Louis-Jeantet Foundation. We would like to thank J. C. Bulinski for providing p3EGFP–EMTB, K. Weijer for EGFP–ABD and K. Ribbeck for fluorescently labelled Ran.

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Correspondence to Jan Ellenberg.

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Supplementary information

Suppplementary Figure and Video Legends

Text to accompany the below Supplementary Figures and Supplementary Videos. (DOC 27 kb)

Supplementary Figure S1

Effects of nocodazole and latrunculin B on the microtubule and actin cytoskeleton (PDF 685 kb)

Supplementary Figure S2

Low doses of LAB also strongly delay chromosome movement. (PDF 135 kb)

Supplementary Figure S3

DNA coated beads nucleate actin patches similar to chromosomes. (PDF 3558 kb)

Supplementary Video S1

Microtubules are too short to capture chromosomes in meiosis I of starfish oocytes. (MOV 3182 kb)

Supplementary Video S2

Chromosome congression in an untreated oocyte. (MOV 3124 kb)

Supplementary Video S3

Chromosome congression in an oocyte preincubated for 1h and matured in 3.3 µM nocodazole. (MOV 2188 kb)

Supplementary Video S4

Chromosome congression in an oocyte treated with 2 µM LAB 10 minutes before NEBD. (MOV 3573 kb)

Supplementary Video S5

Chromosome congression in an oocyte treated with 3.3 µM nocodazole and 250 nM LAB at the time of hormone addition. (MOV 4145 kb)

Supplementary Video S6

A contractile actin meshwork moves chromosomes to the animal pole in starfish oocytes. (MOV 5273 kb)

Supplementary Video S7

Details of the actin mesh. (MOV 2437 kb)

Supplementary Video S8

Phalloidin injection delays the collapse of the actin mesh. (MOV 1978 kb)

Supplementary Video S9

Chromosome congression in an oocyte treated with 120 nM LAB 10 minutes before NEBD. (MOV 1696 kb)

Supplementary Video S10

DNA coated beads nucleate actin patches similar to chromosomes. (MOV 1215 kb)

Supplementary Video S11

Uncoated beads do not nucleate actin. (MOV 2672 kb)

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Lénárt, P., Bacher, C., Daigle, N. et al. A contractile nuclear actin network drives chromosome congression in oocytes. Nature 436, 812–818 (2005) doi:10.1038/nature03810

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