MICROMANIPULATION STUDIES OF CHROMOSOME MOVEMENT .2. BIREFRINGENT CHROMOSOMAL FIBERS AND THE MECHANICAL ATTACHMENT OF CHROMOSOMES TO THE SPINDLE

The degree of mechanical coupling of chromosomes to the spindles of Nephrotoma ferruzinea and Trimeratropis maritima primary spermatocytes varies with the stage of meiosis and the birefringent retardation of the chromosomal fibers. In early prometaphase, before briefringent chromosomal fibers have formed, a bivalent can be displaced toward a spindle pole by a single, continuous pull with a microneedle. Resistance to poleward displacement increases with increased development of the chromosomal fibers, reaching a maximum at metaphase. At this stage kinetochores cannot be displaced > 1 .mu.m toward either spindle pole, even by a force which is sufficient to displace the entire spindle within the cell. The abolition of briefringence with either colcemid or vinblastine results in the loss of chromosome-spindle attachment. In the absence of birefringent fibers a chromosome can be displaced anywhere within the cell. The photochemical inactivation of colcemid by irradiation with 366-nm light results in the reformation of birefringent chromosomal fibers and the concomitant re-establishment of chromosome attachment to the spindle. The hypothesis that the birefringent chromosomal fibers anchor the chromosomes to the spindle and transmit the force for anaphase chromosome movement is supported.