Xenopus tropicalis egg extracts provide insight into scaling of the mitotic spindle.
The African clawed frog Xenopus laevis has been instrumental to investigations of both development and cell biology, but the utility of this model organism for genetic and proteomic studies is limited by its long generation time and unsequenced pseudotetraploid genome. Xenopus tropicalis, which is a small, faster-breeding relative of X. laevis, has recently been adopted for research in developmental genetics and functional genomics, and has been chosen for genome sequencing. We show that X. tropicalis egg extracts reconstitute the fundamental cell cycle events of nuclear formation and bipolar spindle assembly around exogenously added sperm nuclei. Interestingly, X. tropicalis spindles were approximately 30% shorter than X. laevis spindles, and mixing experiments revealed a dynamic, dose-dependent regulation of spindle size by cytoplasmic factors. Measurements of microtubule dynamics revealed that microtubules polymerized slower in X. tropicalis extracts compared to X. laevis, but that this difference is unlikely to account for differences in spindle size. Thus, in addition to expanding the range of developmental and cell biological experiments, the use of X. tropicalis provides novel insight into the complex mechanisms that govern spindle morphogenesis.
Brown
Katherine S
KS
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.
Blower
Michael D
MD
Maresca
Thomas J
TJ
Grammer
Timothy C
TC
Harland
Richard M
RM
Heald
Rebecca
R
D000818:Animals; D002457:Cell Extracts / Q000737:chemistry; D008870:Microtubules / Q000378:metabolism / Q000648:ultrastructure; D010063:Ovum / Q000737:chemistry* / Q000648:ultrastructure; D008941:Spindle Apparatus / Q000378:metabolism / Q000648:ultrastructure*; D014981:Xenopus
D002457:Cell Extracts