Carbon disulfide induces mitochondria-mediated apoptosis in Sertoli-germ cells coculture

Carbon disulfide (CS2), a common organic solvent, induces a variety of adverse effects in the male reproductive system. In this study, we investigated the cytotoxicity, ultrastructural changes, and potential apoptotic induction mechanisms of CS2 in mixed cultures of Sertoli and germ cells. Sertoli and germ cells were cocultured and treated with CS2 for 24 h. Growth rates were noted, and apoptotic cells were identified by Hoechst 33258 staining. Ultrastructure changes were observed via transmission electron microscopy (TEM). Mitochondrial membrane potential and expressions of apoptosis-related factors (cytochrome c, Bax, Bcl-2, caspase-3 and caspase-9) were examined by JC-1 staining, western blot, and real-time PCR. The results showed that CS2 treatment was associated with reduced growth rates of Sertoli-germ cells. Ultrastructure changes in Sertoli-germ cells treated with CS2 were typical of apoptosis. In addition, CS2 treatment depolarized mitochondrial membrane potential, upregulated Bax levels and downregulated Bcl-2 levels, released cytochrome c from the mitochondrial intermembrane space to the cytosol, and triggered mitochondria-mediated apoptosis. Subsequently, caspase-9 and caspase-3 were activated, resulting in Sertoli-germ cells apoptosis. The above data suggest that CS2 has adverse effect on the viability of Sertoli-germ cells and induces apoptosis through mitochondrial pathway.