Luo Chan, Wang Zhiqiang, Wang Jinling, Yun Feng, Lu Fenghua, Fu Jiayuan, Liu Qingyou, Shi Deshun
State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, 530005, China.
College of Animal Science and Technology, Guangxi University, Nanning, 530005, China.
Sci China Life Sci. 2022 Oct;65(10):2076-2092. doi: 10.1007/s11427-021-2039-6. Epub 2022 Mar 25.
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 H3K9 acetylation and low heterochromatin protein 1α (HP1α) expression. High cloning efficiency BFFs had more H3K9ac regions near to the upstream of glycolysis genes by ChIP-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 qRT-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 H3K9 acetylation, decrease in histone deacetylase activity and HP1α expression, 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.
哺乳动物个体的体细胞克隆效率存在差异，但导致这种差异的机制尚不清楚。在此，我们发现高克隆效率的水牛胎儿成纤维细胞（BFFs）表现出强大的能量代谢、更松散的染色质结构、高H3K9乙酰化水平和低异染色质蛋白1α（HP1α）表达。通过ChIP-seq发现，高克隆效率的BFFs在糖酵解基因上游附近有更多的H3K9ac区域，并且通过ATAC-seq发现与糖酵解基因相关的开放位点更多。通过qRT-PCR还发现这些糖酵解基因在高克隆效率的BFFs中表达也更高。糖酵解的两个关键酶，PDKs和LDH，被证实与BFFs的组蛋白乙酰化和染色质开放性有关。用PS48（PDK1激活剂）处理低克隆效率的BFFs导致细胞内乳酸产生增加和H3K9乙酰化增加，组蛋白脱乙酰酶活性和HP1α表达降低，染色质结构凝聚减少，更多的克隆胚胎发育到囊胚阶段。这些结果表明，水牛体细胞的克隆效率与其糖酵解代谢和染色质结构相关，并且可以通过增加糖酵解代谢来提高。
