Microtubule flux mediates poleward motion of acentric chromosome fragments during meiosis in insect spermatocytes.
We applied a combination of laser microsurgery and quantitative polarization microscopy to study kinetochore-independent forces that act on chromosome arms during meiosis in crane fly spermatocytes. When chromosome arms located within one of the half-spindles during prometa- or metaphase were cut with the laser, the acentric fragments (lacking kinetochores) that were generated moved poleward with velocities similar to those of anaphase chromosomes (approximately 0.5 microm/min). To determine the mechanism underlying this poleward motion of detached arms, we treated spermatocytes with the microtubule-stabilizing drug taxol. Spindles in taxol-treated cells were noticeably short, yet with polarized light, the distribution and densities of microtubules in domains where fragment movement occurred were not different from those in control cells. When acentric fragments were generated in taxol-treated spermatocytes, 22 of 24 fragments failed to exhibit poleward motion, and the two that did move had velocities attenuated by 80% (to approximately 0.1 microm/min). In these cells, taxol did not inhibit the disjunction of chromosomes nor prevent their poleward segregation during anaphase, but the velocity of anaphase was also decreased 80% (approximately 0.1 microm/min) relative to untreated controls. Together, these data reveal that microtubule flux exerts pole-directed forces on chromosome arms during meiosis in crane fly spermatocytes and strongly suggest that the mechanism underlying microtubule flux also is used in the anaphase motion of kinetochores in these cells.
LaFountain
J R
JR
Department of Biological Sciences, University at Buffalo, Buffalo, NY 14260, USA. jrl@acsu.buffalo.edu
Oldenbourg
R
R
Cole
R W
RW
Rieder
C L
CL
D000705:Anaphase; D000818:Animals; D016764:Cell Polarity; D002478:Cells, Cultured; D002875:Chromosomes / Q000187:drug effects / Q000378:metabolism*; D004175:Diptera / Q000166:cytology* / Q000187:drug effects; D005454:Fluorescence Polarization; D008297:Male; D008540:Meiosis; D008677:Metaphase; D008858:Microscopy, Phase-Contrast; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D017239:Paclitaxel / Q000494:pharmacology; D013090:Spermatocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D013997:Time Factors
D017239:Paclitaxel