Ptbp2 controls an alternative splicing network required for cell communication during spermatogenesis [RNA-Seq]

Alternative splicing (AS) plays key roles in the specialization of cell functions in different tissues and stages of development. In spermatogenic cells, the complexity of AS isoforms exceeds that of most whole tissues, but the regulation, dynamics, and functions of AS in spermatogenesis have remained poorly defined. We previously demonstrated that the RNA binding protein Ptbp2 is essential for cell survival during spermatogenesis, however the underlying mechanisms were not explored. To investigate this, we generated paired-end RNA-Seq data from wild type (WT) and Ptbp2 conditional knockout (cKO) testes at postnatal day 25 (p25) using Illumina ScriptSeq™ v2 RNA-Seq library preparation kits and sequenced at the CWRU Sequencing Core. Our resulting analyses demonstrate that Ptbp2 is a critical AS regulator in spermatogenic cells, where it controls a functionally-related network of genes involved in germ cell adhesion and protein trafficking to and from the plasma membrane. In parallel, we identified distinct AS programs in different stages of spermatogenesis, and defined a role for Ptbp2 in stage-specific AS regulation. Collectively, the data provide new insights into the importance of AS in mammalian germ cell development and demonstrate a central role for Ptbp2 in its regulation. Transcriptome profiling of testes in wild type (WT) and Ptbp2-deficient (cKO) mice at postnatal day 25 (p25) by deep sequencing, in duplicate using Illumina HiSeq 2500.

可变剪接（Alternative splicing, AS）在不同组织与发育阶段的细胞功能特化进程中发挥关键调控作用。
在生精细胞中，可变剪接异构体的复杂度远超多数完整组织，但目前学界对精子发生过程中可变剪接的调控机制、动态变化与生物学功能仍缺乏深入认知。
我们此前的研究证实，RNA结合蛋白（RNA binding protein）Ptbp2对精子发生过程中的细胞存活至关重要，但未对其背后的分子机制展开探究。
为探究该科学问题，我们使用Illumina ScriptSeq™ v2 RNA测序（RNA-Seq）文库制备试剂盒，对出生后第25天（p25）的野生型（WT）与Ptbp2条件性敲除（cKO）小鼠睾丸样本构建双端RNA测序（paired-end RNA-Seq）文库，并送至凯斯西储大学测序中心（CWRU Sequencing Core）完成上机测序。
后续分析结果显示，Ptbp2是生精细胞中关键的可变剪接调控因子，可调控一系列参与生殖细胞黏附、细胞膜蛋白质双向转运的功能基因网络。
与此同时，我们在精子发生的不同阶段中鉴定出了特异性的可变剪接调控程序，并明确了Ptbp2在阶段特异性可变剪接调控中的功能角色。
综上，本数据集为可变剪接在哺乳动物生殖细胞发育中的重要性提供了全新的研究视角，并证实了Ptbp2在该调控过程中的核心地位。
本数据集包含通过Illumina HiSeq 2500平台开展双重复深度测序得到的、出生后第25天（p25）野生型（WT）与Ptbp2缺陷型（cKO）小鼠睾丸的转录组分析数据。