Cytosine methylation dynamics during spermatogenesis and post-testicular sperm maturation

We carried out whole genome bisulfite sequencing in seven purified sperm populations, including primary spermatocytes, early and late round spermatids, and mature sperm isolated from the caput, corpus, and cauda epididiymis, and from the vas deferens. Overall design: Testicular sperm populations were isolated by elutriation. Epididymal and vas deferens sperm were obtained by gently squeezing sperm out of the dissected tissue. Sperm were washed several times prior to genomic DNA extraction, and purified gDNA was subject to WGBS.

本研究针对7个纯化的精子群体开展了全基因组亚硫酸氢盐测序（whole genome bisulfite sequencing, WGBS），涵盖初级精母细胞（primary spermatocytes）、早圆形精子细胞与晚圆形精子细胞，以及从附睾头（caput epididymis）、附睾体（corpus epididymis）、附睾尾（cauda epididymis）和输精管（vas deferens）中分离得到的成熟精子。实验整体设计如下：睾丸来源的精子群体通过淘析法（elutriation）分离得到；附睾及输精管来源的成熟精子则通过轻柔挤压解剖后的组织获取。所有精子样本在进行基因组DNA（genomic DNA, gDNA）提取前均经过多次洗涤，纯化获得的gDNA将用于WGBS实验。