Changes in fused cells induced by hvj (sendai virus): redistribution of cytoplasmic organelles and cytoskeletal reorganization.

To understand the relationship between the location of organelles and cellular function, we examined the dynamic state of cytoplasmic organelles and cytoskeleton in polynuclear Ehrlich ascites tumor (EAT) cells fused with hemagglutinating virus of Japan (HVJ; Sendai virus) by confocal laser scanning microscopy. Irregular fused cells gradually became spherical during culture, and nuclei and mitochondria were redistributed in the fused cell; nuclei formed a cluster surrounded by mitochondria. F-actin, vimentin, and microtubules were also reorganized with the redistribution of cell organelles. Further, when the morphological change was inhibited by L4-1, a chlorophyll-like substance derived from silkworm faeces, or pyropheophorbide-a, the arrangement of organelles and cytoskeleton remained disturbed, suggesting that the movement of the cytoskeleton is closely associated with cell shape and the distribution of cytoplasmic organelles.

Changes in fused cells induced by hvj (sendai virus): relationship of morphological changes to endocytosis of the surface membrane.

Fused Ehlrich ascites tumor (EAT) cells induced by hemagglutinating virus of Japan (HVJ; Sendai virus) had an irregular shape, reflecting the shape of cell aggregates before fusion. During subsequent culture, the fused cells gradually took on a spherical form within 60 min. Examination of the fused cells revealed a vigorous endocytosis of the cell membrane during the morphological change. When EAT cells were treated with porphyrin derivatives, and the morphological change to a spherical form was inhibited, endocytosis of fused cells was also suppressed, suggesting that the change is closely associated with endocytotic activity. Further examination with porphyrin derivatives and hydrogen peroxide suggested that the inhibition of morphological change is due to the suppression of endocytosis by active oxygen species produced by these substances. Experiments using an endocytotic inhibitor, methylamine, indicated that endocytosis is essential for the morphological change that occurs in the fused cells.