Abstract
Time-lapse imaging with liquid crystal polarized light (LC-PolScope) and fluorescent speckle microscopy (FSM) enabled this study of spindle microtubules in monoastral spindles that were produced in crane-fly spermatocytes through flattening-induced centrosome displacement. Monoastral spindles are found in several other contexts: after laser ablation of one of a cell's two centrosomes (in the work of Khodjakov et al.), in Drosophila "urchin" mutants (in the works of Heck et al. and of Wilson et al.), in Sciara males (in the works of Fuge and of Metz), and in RNAi variants of Drosophila S2 cells (in the work of Goshima et al.). In all cases, just one pole has a centrosome (the astral pole); the other lacks a centrosome (the anastral pole). Thus, the question: How is the anastral half-spindle, lacking a centrosome, constructed? We learned that monoastral spindles are assembled in two phases: Phase I assembles the astral half-spindle composed of centrosomal microtubules, and Phase II assembles microtubules of the anastral half through extension of new microtubule polymerization outward from the spindle's equatorial mid-zone. That process uses plus ends of existing centrosomal microtubules as guiding templates to assemble anastral microtubules of opposite polarity. Anastral microtubules slide outward with their minus ends leading, thereby establishing proper bipolarity just like in normal biastral spindles that have two centrosomes.