Small RNA expression analysis in histone replacement-completed sperm

Increasing evidence indicates that paternal dietary conditions influence the metabolic disorders of offspring. Previous studies suggested the involvement of epigenetic regulation for such phenomena, but the mechanism remains elusive. To reveal the molecular function of stress-dependent epigenetic regulator ATF7 in paternal inheritance of dietry effect on offpring phenotype, we analyzed ATF7-binding profiles in testicular germ cells by ChIP-seq, DNA methylation profiles in sperm by TGBS, small RNA expression profiles in HRCS by RNA-seq,and whole expression profiles in offspring liver by expression array analysis, using wild-type and ATF7 heterozygous mutant mice fed with control diet or low-protein diet. Quantification of small RNA expression in histone replacement-completed sperm (HRCS) collected from four groups of mice; WT_CD, WT_LPD, ATF7+/-_CD and ATF7+/-_LPD.越来越多的证据表明，父代饮食状况可对子代的代谢紊乱产生影响。既往研究提示此类现象与表观遗传调控相关，但具体机制仍未阐明。为揭示应激依赖性表观遗传调控因子ATF7在父代饮食效应影响子代表型的遗传过程中的分子功能，本研究利用喂食正常对照饮食（control diet, CD）或低蛋白饮食（low-protein diet, LPD）的野生型及ATF7杂合突变小鼠，通过染色质免疫沉淀测序（ChIP-seq）分析睾丸生殖细胞中ATF7的结合谱，通过TGBS分析精子中的DNA甲基化谱，通过RNA测序（RNA-seq）分析完成组蛋白替换的精子（histone replacement-completed sperm, HRCS）中的小RNA表达谱，并通过表达芯片分析子代肝脏中的全基因表达谱。此外，本研究对取自四组小鼠——WT_CD、WT_LPD、ATF7+/-_CD及ATF7+/-_LPD——的完成组蛋白替换的精子（HRCS）中的小RNA表达水平进行了定量检测。