Sperm microRNA Content Is Altered in a Mouse Model of Male Obesity, but the Same Suite of microRNAs Are Not Altered in Offspring's Sperm

Abstract

The prevalence of obesity is increasing worldwide and has tripled in men of reproductive age since the 1970s. Concerningly, obesity is not only comorbid with other chronic diseases, but there is mounting evidence that it increases the non-communicable disease load in their children (eg mortality, obesity, autism). Animal studies have demonstrated that paternal obesity increases the risk of metabolic (eg glucose metabolism defects, obesity) and reproductive disorders in offspring. Epigenetic changes within sperm are clear mechanistic candidates that are associated with both changes to the father's environment and offspring phenotype. Specifically there is emerging evidence that a father's sperm microRNA content both responds to paternal environmental cues and alters the gene expression profile and subsequent development of the early embryo. We used a mouse model of high fat diet (HFD) induced obesity to investigate whether male obesity could modulate sperm microRNA content. We also investigated whether this alteration to a father's sperm microRNA content lead to a similar change in the sperm of male offspring. Our investigations were initially guided by a Taqman PCR array, which indicated the differential abundance of 28 sperm borne microRNAs in HFD mice. qPCR confirmation in a much larger cohort of founder males demonstrated that 13 of these microRNAs were differentially abundant (11 up-regulated; 2 down-regulated) due to HFD feeding. Despite metabolic and reproductive phenotypes also being observed in grand-offspring fathered via the male offspring lineage, there was no evidence that any of the 13 microRNAs were also dysregulated in male offspring sperm. This was presumably due to the variation seen within both groups of offspring and suggests other mechanisms might act between offspring and grand-offspring. Thus 13 sperm borne microRNAs are modulated by a father's HFD and the presumed transfer of this altered microRNA payload to the embryo at fertilisation potentially acts to alter the embryonic molecular makeup post-fertilisation, altering its growth trajectory, ultimately affecting adult offspring phenotype and may contribute to paternal programming.

Fig 1. The abundance of the 13 sperm borne microRNAs by qPCR in the sperm of the (A) CD or HFD fed fathers and the (B) male offspring born to by either CD or HFD fed fathers.

Fold change (FC); vs (A) CD fathers or (B) offspring born to CD fathers by ΔΔCt method) of each microRNA is given for the (A) F0 males fed the CD (n = 13) or HFD (n = 14; from 3 separate cohorts) and the (B) F1 males born to either CD (n = 9; black bars) or HFD (n = 9; white bars) fathers. The (A) geometric mean of mmu-miR-10a-5p and mmu-miR-195-5p or (B) mmu-miR-10a-5p alone was used as a reference microRNA (the least variable microRNA(s) across all samples in the present experimental setup). p values denoted [* p < 0.05; **p < 0.01] are derived from (A) univariate general linear modelling or (B) no significant differences were detected by a Student’s T test. Data is presented a means ± SEM.