Journal of Andrology
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Published-Ahead-of-Print December 27, 2006, DOI:10.2164/jandrol.106.000455

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Cystoskeleton Vimentin Disruption of Mouse Sertoli Cells Injured by Nitrogen Mustard in vitro

Dawei He *, Deying Zhang , Guanghui Wei , Tao Lin , and Xuliang Li

* To whom correspondence should be addressed. E-mail: babyuro{at}163.com.

Reproductive toxicity is one of the potential side effects of anticancer alkylating agents, with potential affects on vimentin intermediate filaments, one of the main components of the Sertoli cytoskeleton. Research suggests (Aumuller et al., 1988; Aumuller et al., 1992) that the highly organized and active Sertoli cytoskeleton is important in spermatogenesis. The aim of the current study was to investigate the effects of alkylating agents on vimentin filament expression in vitro. Sertoli cells, isolated from 20-day-old mice testes, were cultured for five days then incubated with 0, 50, 100 and 200 µmol/L nitrogen mustard (HN2). Morphologic changes in Sertoli cells were observed per interval 30 minutes at 12h time points to HN2 100 µmol/L. Vimentin expression was investigated by immunocytochemistry at 6h and 24h post-treatment and RT-PCR and Western blot at 12h post-treatment with 50, 100 and 200 µmol/L HN2. Exposure to HN2 resulted in a comparatively small Sertoli cell body with diminished cytoplasm. Sertoli cells were shrunk or detached. Cytoskeletal disruption increased with increasing HN2 concentrations. The optical density values of vimentin antibody and expression of vimentin mRNA and protein were significantly decreased with increasing concentration of HN2. Significant treatment dose and time-dependent differences of vimentin mRNA and protein expression levels were also noted. Our data suggest that the change in the biochemical properties of vimentin may represent one of mechanism the mechanisms of reproductive toxicity resulting from HN2 is disruption of Sertoli cell vimentin filament structure, accompanied by a down regulation of vimentin expression.


Key words: Testis • Sertoli cell • alkylating agent • vimentin




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