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© The Rockefeller University Press, 0021-9525/1997//1321 $5.00
The Journal of Cell Biology, Volume 137, Number 6, , 1997 1321-1336

Article

Spindle Dynamics during Meiosis in Drosophila Oocytes



Sharyn A. Endow and Donald J. Komma

Department of Microbiology, Duke University Medical Center, Durham, North Carolina 27710

Mature oocytes of Drosophila are arrested in metaphase of meiosis I. Upon activation by ovulation or fertilization, oocytes undergo a series of rapid changes that have not been directly visualized previously. We report here the use of the Nonclaret disjunctional (Ncd) microtubule motor protein fused to the green fluorescent protein (GFP) to monitor changes in the meiotic spindle of live oocytes after activation in vitro. Meiotic spindles of metaphase-arrested oocytes are relatively stable, however, meiotic spindles of in vitro–activated oocytes are highly dynamic: the spindles elongate, rotate around their long axis, and undergo an acute pivoting movement to reorient perpendicular to the oocyte surface. Many oocytes spontaneously complete the meiotic divisions, permitting visualization of progression from meiosis I to II. The movements of the spindle after oocyte activation provide new information about the dynamic changes in the spindle that occur upon re-entry into meiosis and completion of the meiotic divisions. Spindles in live oocytes mutant for a lossof-function ncd allele fused to gfp were also imaged. The genesis of spindle defects in the live mutant oocytes provides new insights into the mechanism of Ncd function in the spindle during the meiotic divisions.

1. Abbreviations used in this paper: blo, bloated; cand, claret nondisjunctional; GFP, green fluorescent protein; Ncd, Nonclaret disjunctional; so, sine oculis; y, yellow.

Special thanks to T. Hazelrigg for gfp–exu DNA, a host stock for P element transformation, and helpful comments, and R. Tsien for the S65 -> T mutant gfp. O.T. Hardy performed the in situ hybridization experiments to map the ncd–gfp and ncd–gfp* insertions. We also thank W. Theurkauf for a protocol for preparation of nonactivated oocytes, W. Sullivan for a generous gift of rhodamine-conjugated anti–{alpha}-tubulin antibody, and W. Rasband for modification of NIH Image. The initial imaging of Ncd–GFP in oocytes was done with T. Salmon, who provided valuable guidance and suggestions.

Please address all correspondence to Sharyn A. Endow, Department of Microbiology, Duke University Medical Center, Durham, NC 27710. Tel.: (919) 684-4311; Fax: (919) 684-8735; E-mail: endow{at}galactose.mc.duke.edu


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