Three-dimensional analysis of mitosis and cytokinesis in the binucleate parasite Giardia intestinalis.
In the binucleate parasite Giardia intestinalis, two diploid nuclei and essential cytoskeletal structures including eight flagella are duplicated and partitioned into two daughter cells during cell division. The mechanisms of mitosis and cytokinesis in the binucleate parasite Giardia are poorly resolved, yet have important implications for the maintenance of genetic heterozygosity. To articulate the mechanism of mitosis and the plane of cell division, we used three-dimensional deconvolution microscopy of each stage of mitosis to monitor the spatial relationships of conserved cytological markers to the mitotic spindles, the centromeres and the spindle poles. Using both light- and transmission electron microscopy, we determined that Giardia has a semi-open mitosis with two extranuclear spindles that access chromatin through polar openings in the nuclear membranes. In prophase, the nuclei migrate to the cell midline, followed by lateral chromosome segregation in anaphase. Taxol treatment results in lagging chromosomes and half-spindles. Our analysis supports a nuclear migration model of mitosis with lateral chromosome segregation in the left-right axis and cytokinesis along the longitudinal plane (perpendicular to the spindles), ensuring that each daughter inherits one copy of each parental nucleus with mirror image symmetry. Fluorescence in situ hybridization (FISH) to an episomal plasmid confirms that the nuclei remain separate and are inherited with mirror image symmetry.
Sagolla
Meredith S
MS
Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, CA 94720, USA.
Dawson
Scott C
SC
Mancuso
Joel J
JJ
Cande
W Zacheus
WZ
D000818:Animals; D002478:Cells, Cultured; D020090:Chromosome Segregation; D048749:Cytokinesis; D005407:Flagella / Q000235:genetics; D016829:Giardia lamblia / Q000235:genetics / Q000648:ultrastructure*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis; D008954:Models, Biological; D010957:Plasmids / Q000235:genetics; D008941:Spindle Apparatus / Q000378:metabolism