Mammalian individuals differ in their somatic cell cloning efficiency,but the mechanisms leading to this variation is poorly understood.Here we found that high cloning efficiency buffalo fetal fibroblasts(BFFs)display...Mammalian individuals differ in their somatic cell cloning efficiency,but the mechanisms leading to this variation is poorly understood.Here we found that high cloning efficiency buffalo fetal fibroblasts(BFFs)displayed robust energy metabolism,looser chromatin structure,high H3 K9 acetylation and low heterochromatin protein 1α(HP1α)expression.High cloning efficiency BFFs had more H3 K9 ac regions near to the upstream of glycolysis genes by Ch IP-seq,and involved more openness loci related to glycolysis genes through ATAC-seq.The expression of these glycolysis genes was also found to be higher in high cloning efficiency BFFs by q RT-PCR.Two key enzymes of glycolysis,PDKs and LDH,were confirmed to be associated with histone acetylation and chromatin openness of BFFs.Treatment of low cloning efficiency BFFs with PS48(activator of PDK1)resulted in an increase in the intracellular lactate production and H3 K9 acetylation,decrease in histone deacetylase activity and HP1αexpression,less condensed chromatin structure and more cloning embryos developing to blastocysts.These results indicate that the cloning efficiency of buffalo somatic cells is associated with their glycolytic metabolism and chromatin structure,and can be improved by increasing glycolytic metabolism.展开更多
基金supported by the National Natural Science Foundation of China(31772597,31972996,31902125)Guangxi Natural Science Foundation(2017GXNSFAA198311)。
文摘Mammalian individuals differ in their somatic cell cloning efficiency,but the mechanisms leading to this variation is poorly understood.Here we found that high cloning efficiency buffalo fetal fibroblasts(BFFs)displayed robust energy metabolism,looser chromatin structure,high H3 K9 acetylation and low heterochromatin protein 1α(HP1α)expression.High cloning efficiency BFFs had more H3 K9 ac regions near to the upstream of glycolysis genes by Ch IP-seq,and involved more openness loci related to glycolysis genes through ATAC-seq.The expression of these glycolysis genes was also found to be higher in high cloning efficiency BFFs by q RT-PCR.Two key enzymes of glycolysis,PDKs and LDH,were confirmed to be associated with histone acetylation and chromatin openness of BFFs.Treatment of low cloning efficiency BFFs with PS48(activator of PDK1)resulted in an increase in the intracellular lactate production and H3 K9 acetylation,decrease in histone deacetylase activity and HP1αexpression,less condensed chromatin structure and more cloning embryos developing to blastocysts.These results indicate that the cloning efficiency of buffalo somatic cells is associated with their glycolytic metabolism and chromatin structure,and can be improved by increasing glycolytic metabolism.
