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  • 1
    ISSN: 1615-6102
    Schlagwort(e): Brilliant sulfoflavin ; Cyanidioschyzon merolae ; Fenton reaction ; Fluorescence microscopy ; Hydrogen peroxide ; Microbody
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary A novel procedure is described for fluorescence staining of microbodies, which can be applied quickly and easily. We developed this technique of microbody staining with the unicellular red algaCyanidioschyzon merolae. Cyanidioschyzon merolae only contains a single chloroplast, mitochondrion, and microbody per cell, and the mitotic cycle and the organelle division cycle are easily synchronized. Knowing that the concentration of H2O2 in the microbody is higher than it is in the cytosol and other cell components, we attempted to visualize the microbody by using fluorescence microscopy to detect H2O2. Brilliant sulfoflavin (BSF), used for detecting Fe2+ in analytical chemistry, fluoresces when it reacts with Fe2+ and H2C2. We were able to specifically stain microbodies with BSF, under acidic conditions (pH 3.0 or pH 2.5) with blue-light excitation. Using this procedure, we observed division of the microbody and the effect of aphidicolin on the microbody. We also discovered that microbody division is regulated by the cell nucleus and follows division of the cell nucleus.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    Digitale Medien
    Digitale Medien
    Springer
    Protoplasma 196 (1997), S. 135-141 
    ISSN: 1615-6102
    Schlagwort(e): Aphidicolin ; Cell cycle ; Checkpoint control ; Cyanidioschyzon merolae ; Microbody ; Mitochondrial division
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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