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Published online June 23, 2008
doi:10.1083/jcb.200712137
The Journal of Cell Biology, Vol. 181, No. 7, 1047-1054
The Rockefeller University Press, 0021-9525 $30.00
© 2008 Goss et al.
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 Both daughter cells traffic and exocytose membrane at the cleavage furrow during mammalian cytokinesis

John W. Goss and Derek K. Toomre

Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510

Correspondence to Derek K. Toomre: derek.toomre{at}yale.edu

Membrane trafficking during cytokinesis is not well understood. We used advanced live cell imaging techniques to track exocytosis of single vesicles to determine whether constitutively exocytosed membrane is focally delivered to the cleavage furrow. Ultrasensitive three-dimensional confocal time-lapse imaging of the temperature-sensitive membrane cargo protein vesicular stomatitis virus protein–yellow fluorescent protein revealed that vesicles from both daughter cells traffic out of the Golgi and into the furrow, following curvilinear paths. Immunolocalization and photobleaching experiments indicate that individual vesicles accumulate at the midbody and generate a reserve vesicle pool that is distinct from endosomal and lysosomal compartments. Total internal reflection fluorescence microscopy imaging provided direct evidence that Golgi-derived vesicles from both daughter cells not only traffic to the furrow region but dock and fuse there, supporting a symmetrically polarized exocytic delivery model. In contrast, quantitative analysis of midbody abscission showed inheritance of the midbody remnant by one daughter cell, indicating that cytokinesis is composed of both symmetrical and asymmetrical stages.

Abbreviations used in this paper: DIC, differential interference contrast; EMMA, exocytic midzone membrane accumulation; GPI, glycosylphosphatidylinositol; ROI, region of interest; SDCM, spinning disc confocal microscopy; TGN, trans-Golgi network; TIRFM, total internal reflection fluorescence microscopy; VSVG, vesicular stomatitis virus glycoprotein.


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