[A fluorescent-cytochemical method for selective detection of yeast mitochondrial DNA].
Yeast mitochondrial DNA (mDNA) can selectively be detected using a specific dye (DAPI). Nuclear DNA (nDNA) was stained along with mDNA only in three out of the fifteen studied yeast strains. Visualisation with a luminescent microscope showed that mDNA content varied among different yeast species as well as the size and shape of fluorescent mitochondrial nuclei. Intensive nDNA fluorescence was detected when a fixed specimen was treated with DAPI. Under these conditions, mDNA was revealed only in yeast cells with its high content. The process of fixation was shown to interfere with the integrity of mitochondria. It is also possible that the structure of DNA was modified to affect its interaction with the dye and thus the level of fluorescence. The developed technique of selective mDNA staining and the experimental results make it possible to gain a more accurate quantitative information about DNA content at the cellular level using cyto- and spectrofluorimetric techniques. This study involves important aspects pertinent to the dye interaction with yeast nuclear and mitochondrial DNA, both native and subjected to different fixation procedures.
Poglazova
M N
MN
Mashkovtseva
A V
AV
D004271:DNA, Fungal / Q000032:analysis*; D004272:DNA, Mitochondrial / Q000032:analysis*; D005456:Fluorescent Dyes; D005470:Fluorometry; D007211:Indoles; D008856:Microscopy, Fluorescence; D013050:Spectrometry, Fluorescence; D015003:Yeasts / Q000235:genetics*
D004271:DNA, Fungal; D004272:DNA, Mitochondrial; D005456:Fluorescent Dyes; D007211:Indoles; C007293:DAPI
Fluorestsentno-tsitokhimicheskiĭ metod izbiratel'nogo vyiavleniia mitokhondrial'noĭ DNK drozhzheĭ.