Nanopore sequencing reveals the roles of spermatozoal DNA N6-Methyladenine modification in mediating transgenerational lipid metabolism disorder induced by folic acid excessive consumption in chicken [RNA-seq]

Increased consumption of folic acid is prevalent due to its beneficial effects to help protecting from genome damage. Growing evidence is emphasizing the side effects pointing to excessive dietary intake. Effects of paternal folic acid excessive consumption on the offspring and its transgenerational inheritance mechanism remain elusive. We hypothesize that the folic acid excessive consumption will alter sperm DNA N6-methyladenine (6mA) and 5-methylcytosine (5mC) methylation, and heritably influence offspring metabolic homeostasis. This is the results of mRNA sequencing part Overall design: Here, we fed roosters either folic acid-control or folic acid-excess diet throughout life. Paternal chronic folic acid excessive supplementation increased hepatic lipogenesis and lipid accumulation, but reduced lipolysis both in the roosters and their offspring, which was further confirmed to be induced by one carbon metabolism inhibition and gene expression alteration associated with PARR pathway. By nanopore sequencing, the spermatozoal genome-wide DNA (6mA and 5mC) methylation analysis revealed differential methylation pattern of genes implicated in lipid metabolic process. Notably, sperm DNA 6mA methylation could be involved in regulating lipid metabolism related genes expression in offspring chickens.