Microcystin-leucine-arginine affects brain gene expression programs and behaviors of offspring through paternal epigenetic information

Microcystin-leucine-arginine (MC-LR) adversely affects male reproduction and interferes with the development of the offspring. Here, we establish a zebrafish (Danio rerio) model to understand the cross-generational effects of MC-LR in a male-lineage transmission pattern. F0 embryos were reared in water containing MC-LR (0, 5, and 25 μg/L) for 90 days and the developmental indices of F1 and F2 embryos were then measured with no MC-LR treatment. The results show that paternal MC-LR exposure reduced the hatching rate, heart rate and body weight in F1 and F2 generations. Global DNA methylation significantly increased in sperm and testes with the elevation expressions of DNA methyltransferases. Meanwhile, DNA methylation of brain-derived neurotrophic factor (bdnf) promoter was increased in sperm after paternal MC-LR exposure. Subsequently, increased DNA methylation of bdnf promoter and decreased gene expression of bdnf in the brain of F1 male zebrafish were detected. F1 offspring born to F0 males exhibit the depression of BDNF/AKT/CREB pathway and recapitulate these paternal neurodevelopment phenotypes in F2 offspring. In addition, the DNA methylations of dio3b and gad1b promoters were decreased and gene expressions of gad1b and dio3b were increased, accompanied with neurotransmitter disturbances in the brain of F1 male zebrafish after paternal MC-LR exposure. These data revealed that MC-LR displays a potential epigenetic impact on the germ line, reprogramming the epigenetic and transcriptional regulation of brain development, and contributing to aberrant expression of neurodevelopment-related genes and behavior disorders.