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  • 1
    ISSN: 1615-6102
    Keywords: Mitochondrial nuclei ; Mitochondrial division ; Mitochondrial replicon cluster ; Physarum polycephalum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We investigated the manner of mitochondrial DNA (mtDNA) replication and distribution during the culture ofPhysarum polycephalum amoebae cells by microphotometry, anti-BrdU immunofluorescence microscopy, and quantitative hybridization analysis. In amoebae cells ofP. polycephalum, the number of mitochondria per cell and the shape of both mitochondria and mitochondrial nuclei (mt-nuclei) noticeably changed over the culture period. At the time of transfer, about 27 short ellipsoidal shaped mitochondria, which each contained a small amount of DNA, were observed in each cell. The number of mitochondria per cell decreased gradually, while the amount of mtDNA in an mt-nucleus and the length of mt-nuclei increased gradually. Midway through the middle logarithmic growth phase, the number of mitochondria per cell reached a minimum (about 10 mitochondria per cell), but most mtnuclei assumed an elongated shape and contained a large amount of mtDNA. During the late log- and stationary-growth phase, the number of mitochondria per cell increased gradually, while the amount of DNA in an mt-nucleus and mt-nuclei length decreased gradually. Upon completion of the stationary phase, the number and condition of mitochondria within cells returned to that first observed at the time of transfer. The total amount of mtDNA in a cell increased about 1.6-fold the first day, decreased immediately, then maintained a constant level ranging from 130 to 160 T. Except for the fact that mtDNA synthesis began earlier than synthesis of cell nuclei, the rate of increase in mtDNA paralleled that of cell-nuclear DNA throughout the culture. These results indicate that mtDNA is continuously replicated in pace with cell proliferation and the rate of mitochondrial division varies during culture; this mitochondrial division does not synchronize with either mtDNA replication or cell division. Furthermore, we observed the spatial distribution of DNA replication sites along mt-nuclei. Replication began at several sites scattered along an mt-nucleus, and the number of replication sites increased as the length of mt-nuclei increased. These results indicate that mtDNA replication progresses in adjacent replicons, which are collectively termed a mitochondrial replicon cluster.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1615-6102
    Keywords: Mitochondrion-dividing ring ; Plastid-dividing ring ; Mitochondrial division ; Plastid division ; Microbody ; Cyanidioschyzon merolae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We present strong electron microscopic evidence that the mitochondrial-dividing ring (MD-ring) forms as a closed ring about 50 nm wide and 10 nm thick, at the contact point where the micro-body attaches to the mitochondrion. This ring forms in the cytoplasm around an equatorial plane perpendicular to the major axis of a mitochondrion. As the MD-ring increases in both width and thickness, the mitochondrion becomes dumbbell-shaped with a narrow interconnecting isthmus. Then, by successive contractions of the ring, the dumbbell-shaped mitochondrion separates to generate two daughter mitochondria. We also observed formation of an electron dense plastid-dividing ring (PD-ring) during plastid divisions. We noted too the behaviour of the MB in relation to the contraction of MD-rings and PD-rings.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 196 (1997), S. 135-141 
    ISSN: 1615-6102
    Keywords: Aphidicolin ; Cell cycle ; Checkpoint control ; Cyanidioschyzon merolae ; Microbody ; Mitochondrial division
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary It is generally accepted that mitochondria proliferate by division. However, since the apparatus for mitochondrial division was discovered only recently, the basic mechanism of mitochondrial division remains poorly understood. The unicellular red algaCyanidioschyzon merolae is the only organism in which the existence of the apparatus for mitochondrial division (mitochondrion-dividing ring) has been proved by electron microscopy. Since mitochondrial division, mitosis, and cytokinesis regularly occurred in that order, we can assume that tight linkage exists between mitochondrial division and the mitotic cycle. To examine this assumption, we performed experiments with aphidicolin, a specific inhibitor of DNA polymerase α, using cells that had been synchronized by a 12 h light/12 h dark treatment. The effects of aphidicolin onC. merolae cells were examined by both epifluorescence and electron microscopy. When cells synchronized at the S phase were treated with aphidicolin, neither mitosis nor cytokinesis occurred. Epifluorescence microscopy after staining with 3,3′-dihexyloxacarbocyanine iodide (DiOC6; a mitochondrion-specific fluorochrome) revealed that mitochondrial division was also completely inhibited. Nevertheless, electron-microscopic examination of the aphidicolin-treated cells clearly revealed the presence of a mitochondrion-dividing ring in mitochondria in all cells examined, in spite of the absence of mitochondrial division. Microbodies, which might be related to mitochondrial division inC. merolae, also failed to divide and became attached to the mitochondrion-dividing rings. These results imply the presence of a checkpoint control mechanism that inhibits division of mitochondria and microbodies in the absence of the synthesis of cell-nuclear DNA.
    Type of Medium: Electronic Resource
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