Changes in physicochemical properties of microtubules lead to the formation of a single spherical structure of mitochondrial assembly enveloping nuclear chromatins

Treatment of 143B cells with microtubule-active drugs (MADs) including taxol, nocodazole and colchicine induced distinct structural changes, such as rounding of the cells with perinuclear clustering of mitochondria, when the cells were treated for up to 10 h. When the incubation time with MADs was longer than 10 h, multinuclear cells appeared, and their population increased with time. In this study perinuclear clustering of mitochondria i.e. mitochondria encircling the aggregated chromatin of the nucleus that had lost the nuclear membrane was detected. This observation was distinct from that reported in the literature. Mitochondria were aligned in a few lines; the occurrence of mitochondria in even a single line is an extreme case, resulting in one plane of section for electron microscopy. Three-dimensional reconstructions of confocal microscopic images of mitochondria revealed that they were assembled as a spherical structure. The majority of the cells with perinuclear clustering of mitochondria remained intact for up to 24 h. Mitochondria were observed to be clustered around the nucleus in the orthodox configuration or in some cases they were moderately condensed, as observed electron microscopically. Annexin V and PI double staining of cells showed that more than 90% of cells were viable. In the case of treatment with taxol, membrane potential of mitochondria per cell was well maintained although it was moderately lowered in the case of treatment with nocodazole. Taking into consideration the previous data reported from our laboratory, the present results may assist in elucidation of the behaviour of mitochondria during the dividing processes of mammalian cells, which is yet to be clarified.