Molecular alterations underlying the enhanced disruption of spermatogenesis by 2,5-hexanedione and carbendazim co-exposure

The current study investigated the co-exposure effects of 2,5-hexanedione (HD) and carbendazim (CBZ) on gene expression underlying the enhanced pathology previously observed. Adult male rats were exposed to HD (0.33 or 1%) followed by CBZ (67 or 200 mg/kg), and testis samples were collected after 3 and 24 h. Microarray analysis at 3 h revealed that CBZ and HD interact in an agonistic, or synergistic, way at the gene level. Further analysis of candidate genes by qRT-PCR at both 3 and 24 h after co-exposure, revealed that Loxl1 and Clca2/Clca4l were both decreased in expression. Immunohistochemical analysis of Loxl1 at 24 h revealed that Loxl1 is localized to the seminiferous tubules, with the most intense staining in the basement membrane, blood vessels, and acrosomes, with the relative intensity reflecting the gene level changes at 3 h. These findings provide candidate genes for further investigation of the testicular response to damage.

► CBZ and HD interact in a synergistic way at both the histological level and at the gene level.
► Combined exposure to HD and CBZ has synergistic effects on the expression of specific genes, including Clca2/Clca4l and Loxl1.
► The localization of Loxl1 within the testis has not been previously investigated and Loxl1 was found to be localized to several areas within the seminiferous tubules with the most intense staining localized to the acrosomes.