Multigenerational paternal high-fat diet gradually exacerbates environmental stress-induced testicular dysregulated expression of mRNAs in offspring

To investigate the mechanism by which paternal exposure to high-fat diet (HFD) exacerbates environmental stress-impaired testicular germ cell development in offspring, testicular RNA sequencing was performed in H+Cd (the offspring were treated with Cd after paternal exposure to one-generational HFD) and H+H+Cd (the offspring were treated with Cd after paternal exposure to bi-generational HFD) groups. Compared with HFD1D group and HFD2D group, 229 mRNAs were upregulated and 268 mRNAs were downregulated, screened for a 1.2-fold change and adjusted with P < 0.05. GO analysis revealed that downregulated mRNAs were related to multiple biological processes, including the "retinol metabolic process," the "reproductive process," and the "spermatogenesis". F0 generation male mice were fed NC or HFD from 5 weeks to 15 weeks old, and then mated with NC-fed female mice to breed F1 generation. Similarly, partial F1 generation male mice continued to be fed HFD for 10 weeks, and mated with normal female to breed F2 generation. Male mice of F1 and F2 generations were exposed to CdCl2 for 10 weeks, and named H+Cd or H+H+Cd group respectively. Total RNA of mouse testes was extracted, and quality inspections were performed. The sequencing library was constructed with 2.0 μg total RNA from each sample.