RNA sequencing analysis of unilateral orchidectomy, bilateral orchidectomy, and Nell2 knockout initial segment-caput epididymis.

Purpose: The purpose of this study is to characterize genes whose expressions in the initial segment (IS)-caput epididymis were affected by unilateral orchidectomy (UOD), bilateral orchidectomy (BOD), and Nell2 gene ablation. Methods: The unilateral or bilateral orchidectomy of WT mice was performed from 8 weeks old and continued for 4 weeks until tissue sampling at 12 weeks old. Contralateral epididymis was used as a control for UOD. Sham operation was also done as a control for BOD. Nell2-/- mice and their control WT mice were used wthen they were 14-week-old. IS-caput epididymal mRNA profiles were generated by deep sequencing, in triplicate, using Illumina NovaSeq6000. Results: RNA-seq data identified differentially expressed transcripts. Conclusions: Our results show that the expression of many genes was affected by UOD, BOD, and Nell2 ablation. IS-caput epididymal mRNA profiles of mice with experimental treatments were generated by deep sequencing, in triplicate, using Illumina NovaSeq6000.研究目的：本研究旨在鉴定并表征在初始段（initial segment, IS）-附睾头部（caput epididymis）中，其表达受单侧睾丸切除术（unilateral orchidectomy, UOD）、双侧睾丸切除术（bilateral orchidectomy, BOD）以及Nell2基因敲除（Nell2 gene ablation）影响的基因。 实验方法：选取8周龄的野生型（wild type, WT）小鼠实施单侧或双侧睾丸切除术，术后饲养4周，于12周龄时采集组织样本。以对侧附睾作为单侧睾丸切除术的对照，同时设置假手术组作为双侧睾丸切除术的对照。另选取14周龄的Nell2基因敲除（Nell2 gene ablation, Nell2-/-）小鼠及其同窝野生型对照小鼠开展实验。采用Illumina NovaSeq6000平台对IS-caput附睾组织的mRNA进行深度测序，每组设置3次重复，以获取转录组表达谱。 实验结果：通过RNA测序鉴定得到差异表达转录本。 研究结论：本研究结果显示，单侧睾丸切除术、双侧睾丸切除术及Nell2基因敲除均可影响众多基因的表达。经不同实验处理的小鼠IS-caput附睾组织的mRNA转录组表达谱，通过深度测序（采用Illumina NovaSeq6000平台，设置3次重复）得以生成。