Paternal Methyl Donor Supplementation in Rats Improves Fertility, Physiological Outcomes, Gut Microbial Signatures and Epigenetic Markers Altered by High Fat/High Sucrose Diet

Abstract

Increased consumption of high fat/sucrose (HF/S) diets has contributed to rising rates of obesity and its co-morbidities globally, while also negatively impacting male reproductive health. Our objective was to examine whether adding a methyl donor cocktail to paternal HF/S diet (HF/S+M) improves health status in fathers and offspring. From 3–12 weeks of age, male Sprague Dawley rats consumed a HF/S or HF/S+M diet. Offspring were followed until 16 weeks of age. Body composition, metabolic markers, gut microbiota, DNA methyltransferase (DNMT) and microRNA expression were measured in fathers and offspring. Compared to HF/S, paternal HF/S+M diet reduced fat mass in offspring (p < 0.005). HF/S+M fathers consumed 16% fewer kcal/day, which persisted in HF/S+M female offspring and was explained in part by changes in serum glucagon-like peptide-1 (GLP-1) and peptide tyrosine tyrosine (PYY) levels. Compared to HF/S, HF/S+M fathers had a 33% improvement in days until conception and 300% fewer stillbirths. In fathers, adipose tissue DNMT3a and hepatic miR-34a expression were reduced with HF/S+M. Adult male offspring showed upregulated miR-24, -33, -122a and -143 expression while females exhibited downregulated miR-33 expression. Fathers and offspring presented differences in gut microbial signatures. Supplementing a paternal HF/S diet with methyl-donors improved fertility, physiological outcomes, epigenetic and gut microbial signatures intergenerationally.

Effects of paternal high-fat diet and maternal rearing environment on the gut microbiota and behavior

“…We found that these interaction effects were also observed in compositional analyses of microbial beta-diversity, such that paternal high-fat diet drives the presence and absence of keystone species in offspring. Again, like previous studies of paternal and/or maternal HFD feeding 84 , 89 , we show a more subtle effect of paternal HFD on offspring gut microbiomes, similar to paternal prebiotic 85 and sucrose diets 86 . We identified a profound influence of rearing environment, supporting previous evidence that identified early life care and stress as key drivers of offspring microbiome beta-diversity 57 , 90 .…”

“…While we do not replicate the diet-induced differences in diversity, our model excluded maternal HFD-feeding and eliminated HFD metabolites in breast milk, unlike other studies. Other studies with controlled paternal-only feeding have shown similarly limited effects of paternal prebiotic 85 and sucrose diets 86 on alpha diversity in F 1 offspring microbiomes. Interestingly, we identified several significant alterations in alpha diversity based on offspring odor exposure.…”

Effect of Paternal Diet on Spermatogenesis and Offspring Health: Focus on Epigenetics and Interventions with Food Bioactive Compounds

“…A recent study [ 141 ] in rats assessed whether adding a methyl donor cocktail [betaine (5 g/kg diet), choline (5.37 g/kg diet), folic acid (5.5 mg/kg diet), and vitamin B12 (0.5 mg g/kg diet)] to the paternal high fat/sucrose diet improves health status in fathers and offspring. Such paternal intervention before conception reduced energy intake and increased levels of GLP-1 and PYY hormones, which are known to reduce food intake.…”

“…More specifically, the methyl-donor intervention decreased the offspring hepatic expression of miR-34a and increased miR-103, miR-107, and miR-33, which are involved in lipid metabolism and the regulation of insulin sensitivity. It further elevated retroperitoneal adipose tissue expression of DNMT1, DNMT3a, and DNMT3 in the adult offspring [ 141 ]. However, when a similar dietary approach (5 g/kg diet choline chloride, 15 g/kg diet betaine, 7.5 g/kg diet methionine, 15 mg/kg diet folic acid, 1.5 mg/kg diet vitamin B12) was used on male animals on LPD, no protection was observed regarding placental physiology and epigenetic regulation [ 142 ].…”

Paternal high protein diet modulates body composition, insulin sensitivity, epigenetics, and gut microbiota intergenerationally in rats

“…Real-time PCR was performed as previously described 32 using previously published microRNA primer sequences. 33 Relative gene expression was calculated using the 2 -ΔCT method, 34 wherein results were normalized to both SNORD68 and SNORD96A controls genes. We selected a panel of 10 microRNAs whose differential expression in liver tissue has been previously linked with obesity and insulin resistance.…”