Disruption of cytoskeleton by methylmercury in cultured CHO cells.
The effect of methylmercury (MM) on three main cytoskeletal components [i.e. microtubules (MT), microfilaments (MF) and intermediate filaments (IF)] and on specific biochemical parameters (i.e. glutathione transferase (GST), glutathione reductase (RED), glutathione peroxidase (GSH-Px), glyoxalase 1 (GLY 1) and total -SH groups (TSH) of the cytosolic fraction) was studied in cultured Chinese hamster ovary (CHO) cells. The experiments were conducted with increasing doses of MM (i.e. 1, 4 and 8 mum), using an exposure time of 16 hr; and with a fixed dose of MM (2 mum), using increasing exposure periods (i.e. 0-24 hr). The morphological changes observed by immunofluorescence seemed to indicate that MF were damaged as much as (if not more than) MT after 16 hr of exposure to 4 mum-MM. At a concentration of 1 mum, MM only affected MF. The time-course experiments revealed that IF as well as MF and MT were severely disorganized after 3 and 6 hr of incubation in the presence of 2 mum-MM. However, an obvious reorganization was observed after 24 hr of exposure. In experiments using increasing MM doses, changes in the enzymatic activities were less noticeable than those observed in the morphology; only a modest decrease in TSH and RED activities (<30%) was recorded at the highest dose of MM used (i.e. 8 mum). In contrast, increasing the time of exposure to MM induced changes in both the cytoskeletal structures and the biochemical parameters: the lowest RED activity and TSH were observed after 3-6 hr exposure; control values were obtained after an exposure period of 24 hr. Ultrastructural observations on cells treated with increasing doses of MM showed changes in plasmamembrane profile, cytoskeleton organization and mitochondrion structure. The results confirm that MM causes non-specific damage to CHO cells and suggest that a functional interaction may exist between GSH-dependent enzymes and cytoskeletal structures.
Vignani
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R
Department of Environmental Biology, Faculty of Science, University of Siena, Via Mattioli 4, 54100 Siena, Italy.
Milanesi
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C
Di Simplicio
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