Ultraviolet microbeam irradiations of epithelial and spermatocyte spindles suggest that forces act on the kinetochore fibre and are not generated by its disassembly.
Ultraviolet (UV) microbeam irradiations of crane-fly spermatocyte and newt epithelial spindles severed kinetochore fibres (KT-fibres), creating areas of reduced birefringence (ARBs): the remnant KT-fibre consists of two "stubs," a pole-stub attached to the pole and a KT-stub attached to the kinetochore. KT-stubs remained visible but pole-stubs soon became undetectable [Forer et al., 1996]. At metaphase, in both cell types the KT-stub often changed orientation immediately after irradiation and its tip steadily moved poleward. In spermatocytes, the chromosome attached to the KT-stub remained at the equator as the KT-stub elongated. In epithelial cells, the KT-stub sometimes elongated as the associated chromosome remained at the equator; other times the associated chromosome moved poleward together with the KT-stub, albeit only a short distance toward the pole. When an ARB was generated at anaphase, chromosome(s) with a KT-stub often continued to move poleward. In spermatocytes, this movement was accompanied by steady elongation of the KT-stub. In epithelial cells, chromosomes accelerated polewards after irradiation until the KT-stubs reached the pole, after which chromosome movement returned to normal speeds. In some epithelial cells fine birefringent fibres by chance were present along one edge of ARBs; these remnant fibres buckled and broke as the KT-stub and chromosome moved polewards. Similarly, KT-stubs that moved into pole stubs (or astral fibres) caused the pole stubs (or astral fibres) to bend sharply from the point of impact. Our results contradict models of chromosome movement that postulate that force is generated by the kinetochore disassembling the KT-fibre. Instead, these results suggest that poleward directed forces act on the KT-fibre and the KT-stub and suggest that continuity of microtubules between kinetochore and pole is not obligatory for achieving anaphase motion to the pole.
Spurck
T
T
School of Botany, University of Melbourne, Parkville, Vic., Australia.
Forer
A
A
Pickett-Heaps
J
J
D000705:Anaphase; D000818:Animals; D004175:Diptera; D004848:Epithelium / Q000528:radiation effects; D018386:Kinetochores / Q000528:radiation effects* / Q000648:ultrastructure; D008297:Male; D008677:Metaphase; D008938:Mitosis; D012455:Salamandridae; D013090:Spermatocytes / Q000166:cytology / Q000502:physiology / Q000528:radiation effects*; D008941:Spindle Apparatus / Q000502:physiology / Q000528:radiation effects* / Q000648:ultrastructure; D014466:Ultraviolet Rays