Comprehensive analysis of Poly(A) tails in Mouse Testes and Ovaries using Nanopore direct RNA Sequencing.

Germ cell development, gametogenesis, is a complex process in which dysfunction leads to infertility, a growing health and sociological problem worldwide. In both spermatogenesis and oogenesis, post-transcriptional gene expression regulation is crucial. Essentially, all mRNAs possess non-templated poly(A) tails, which composition and dynamics (elongation, shortening, and modifications) determine the fate of mRNA. In this context, we present a comprehensive transcriptomic dataset that focuses on mRNA poly(A) tails metabolism in murine testes. Analyzed RNA samples are isolated from wild-type and transgenic mice lacking TENT5C polymerase, which can extend poly(A) tails in the cytoplasm. Both whole testes material, as well as male gametes at different developmental stages (round or long spermatids) were analyzed. To investigate the role of Tent5c in spermatogenesis we generated RNAseq libraries from (1) whole testes (2) and round or (3) elongated spermatids isolated from either WT or Tent5c -/- mice, followed by differential expression analysis.生殖细胞发育即配子发生是一个复杂的生物学过程，其功能异常会引发不孕症，这已成为全球范围内日益严峻的健康与社会问题。在精子发生与卵子发生过程中，转录后基因表达调控均发挥关键作用。本质上，所有信使RNA（mRNA）均带有非模板依赖型多腺苷酸（poly(A)）尾，其组成与动态变化（延伸、缩短及修饰）直接决定了mRNA的命运。在此研究背景下，本研究公开了一套聚焦于小鼠睾丸中mRNA多腺苷酸尾代谢的全面转录组数据集。本次分析的RNA样本分离自野生型（WT）及缺失TENT5C聚合酶的转基因小鼠——该聚合酶可在细胞质中延伸多腺苷酸尾。研究同时分析了完整睾丸组织，以及不同发育阶段的雄性配子（圆形精子细胞与长形精子细胞）。为探究Tent5c在精子发生过程中的功能，本研究分别从野生型及Tent5c基因敲除（Tent5c -/-）小鼠中分离得到（1）完整睾丸组织、（2）圆形精子细胞以及（3）长形精子细胞，并构建了RNA测序（RNAseq）文库，随后开展差异表达分析。