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Published online January 29, 2007
doi:10.1083/jcb.200605199
The Journal of Cell Biology, Vol. 176, No. 3, 295-305
The Rockefeller University Press, 0021-9525 $30.00
© 2007 Dumont et al.
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Article

 A centriole- and RanGTP-independent spindle assembly pathway in meiosis I of vertebrate oocytes

Julien Dumont1, Sebastian Petri2, Franz Pellegrin1, Marie-Emilie Terret1, Markus T. Bohnsack2, Pascale Rassinier1, Virginie Georget3, Petr Kalab4, Oliver J. Gruss2, and Marie-Hélène Verlhac1

1 UMR7622, Centre National de la Recherche Scientifique/Université Pierre et Marie Curie, 75005 Paris, France
2 Zentrum für Molekulare Biologie der Universität Heidelberg, 69120 Heidelberg, Germany
3 Institut de Biologie Intégrative, IFR83, 75005 Paris, France
4 Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720

Correspondence to Marie-Hélène Verlhac: Marie-Helene.Verlhac{at}snv.jussieu.fr

Spindle formation is essential for stable inheritance of genetic material. Experiments in various systems indicate that Ran GTPase is crucial for meiotic and mitotic spindle assembly. Such an important role for Ran in chromatin-induced spindle assembly was initially demonstrated in Xenopus laevis egg extracts. However, the requirement of RanGTP in living meiotic cells has not been shown. In this study, we used a fluorescence resonance energy transfer probe to measure RanGTP-regulated release of importin ß. A RanGTP-regulated gradient was established during meiosis I and was centered on chromosomes throughout mouse meiotic maturation. Manipulating levels of RanGTP in mice and X. laevis oocytes did not inhibit assembly of functional meiosis I spindles. However, meiosis II spindle assembly did not tolerate changes in the level of RanGTP in both species. These findings suggest that a mechanism common to vertebrates promotes meiosis I spindle formation in the absence of chromatin-induced microtubule production and centriole-based microtubule organizing centers.

M.-E. Terret's present address is Memorial Sloan-Kettering Cancer Center, New York, NY 10021.

M.T. Bohnsack's present address is Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh Eh9 3JR, Scotland, UK.

Abbreviations used in this paper: CSF, cytostatic factor; dbcAMP, dibutiryl cAMP; FRET, fluorescence resonance energy transfer; GV, germinal vesicle; GVBD, GV breakdown; MT, microtubule; MTOC, MT organizing center; RCC1, regulator of chromosome condensation.


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