File name: Nedd5.mov
Authors: Makoto Kinoshita, Tokuko Haraguchi and Yasushi Hiraoka
Playback rate: 8 frames/second
Legend of the movie: This movie shows the dynamic behavior of Nedd5-GFP (green) and chromosomes (red) in living HeLa cells. Nedd5 accumulated near the contractile ring from anaphase through telophase, and finally condensed into the midbody.
Background: Nedd5 is a ubiquitously expressed mammalian septin. The septins comprise a cytoskeletal GTPase subfamily which localize near the cleavage furrow/contractile ring in dividing cells and are essential for the proper budding/cytokinesis in S. cereviciae and D. melanogaster. The live images suggest that Nedd5 plays a similar role in the mammalian cell division.
1. Kinoshita et al., 1997, Genes Dev 11, 1535-1547.
Methods: Transfection; liposome-mediated transfection of plasmid DNA. Microscopy; The DeltaVision (Applied Precision Inc. Seattle, USA) fluorescence microscope system. Microscope equipments; an Olympus inverted microscope IX70, UApo 40x ( NA=1.35), a Peltier-cooled CCD camera (photometrics Ltd., Tucson, USA), a 2 m optical fiber for illumination, a UNIX workstation Indigo2 (Silicon Graphics, USA), a temperature controlled room. The experiment was carried out at 37 C and the time-lapse image data were collected every 1 min at an exposure of 0.5 sec for Nedd5-GFP and 0.5 sec for Hoechst33342.