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Journal of Cell Science, Vol 5, 403-431, Copyright © 1969 by Company of Biologists
Submitted on August 3, 1968
Revised on February 24, 1969
1 Department of Bacteriology
2 Department of Botany, University of Western Ontario London, Ontario, Canada; Department of Genetics, University of Birmingham, P.O. Box 363, Birmingham 15, England.
Mitosis in growing hyphae of haploid and diploid Aspergillus nidulans has been studied with the light microscope in Helly-fixed preparations. At the periphery of the nuleus a sharply defined granule is invariably found which has strong affinity for acid fuchsin. Mitosis begins with the duplication of this granule and the development inside the nucleus of a gradually lengthening fibre between the daughter granules. This fibre is also visibel in dividing nuclei of living hyphae examined by phase-contrast microscopy, when it appears as a grey bar traversing the non-nucleolar region. Division of the chromatin has been studied in preparations stained with aceto orcein, directly or after hydrolysis. It proceeds through four phases, beginning with the condensation of distinct chromosomes from the chromatin network of the resting nuclei. Later the chromosomes become arranged in two parallel rows of chromatinic masses in which individual chromosomes can no longer be distinguished. Division is completed by migration of the chromatin to the ends of each row, giving the appearance of a transverse break in the double bar. The chromatin accumulating at the ends of the double bar condenses to form the two daughter nuclei. The number of chromatinic elements appears to be the same in both haploid and diploid nuclei, although the individual elements are larger in the latter.
Successive staining of the same dividing nucleus, first for the fibre (acid fuchsin) and then for the chromosomes (HCl-aceto orcein or HCl-Giemsa) has established that the fibre lies among the chromosomes, and that is elongation is closely related to the anaphase-telophase movements of the chromosomes. This suggests that the fibre is the equivalent of a mitotic spindle. That the fibre is connected with the chromosomes is further suggested by the observation that, in stained preparations, the fibre is considerably thicker in dividing diploid than in haploid nuclei.
Electron microscopy of dividing nuclei has revealed that mitosis is carried out within the intact nuclear envelope. The mitotic spindle is composed of a bundle of fibrils which traverses the nucleus between two plaques of dense material associated with the envelope.
In order to explain the configurations seen during the division of the chromatin it is proposed that the chromosomes become attached to the central spindle at various points along its length. With the elongation of the spindle the sister chromatids separate and pass, asynchronously, to opposite ends of the spindle along two preferred lines, thereby producing the characteristic double bar figure. The absence of a metaphase plate is thus accounted for by the scattered points of attachment of the chromosomes to the spindle and by their asynchronous division.
Submitted on August 3, 1968
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