RNA Expression Profiling Of Normal Mouse Live Across a Reproductive Cycle

Purpose: To perform RNA-Seq expression profiling of the normal rodent liver at nulliparous, lactation, and involution day 6 reproductive stages. Methods: RNA was extracted and isolated from flash frozen murine liver using the Direct-zol RNA MiniPrep kit. An input of 100ng of RNA was used for library preparation. Library construction was performed by Novogene using a NEBNext® Ultra RNA Library Prep Kit for Illumina. Qualified libraries were sequenced on an Illumina Novaseq6000 Platform using a paired-end 150 run. Results: We identified gene pathway differences bile acid production and FGFR4 signaling between different reproductive stages with upregulation of these pathways during lactation. Additionally, individual genes involved with promoting bile acid synthesis were elevated at lactation whereas genes associated with inhibiting bile acid synthesis were reduced during lactation and elevated during involution. Mouse liver derived mRNA from 7 nulliparous mice, 6 lactation stage mice, and 6 involution day 6 mice generated by 150-cycle paired-end sequencing using the Illumina Novaseq6000 Platform. Fastq files were aligned to mm10 mouse reference genome (GRCm38.89) and per-gene counts quantified by RSEM (version 1.3.1) based on the gene annotation Mus_musculus.GRCm38.89.chr.gtf. Differential gene expression analysis was performed using DESeq2 (version 1.22.2).研究目的：本数据集旨在对处于未产（nulliparous）、泌乳以及泌乳退化第6天三个生殖阶段的正常啮齿类动物肝脏开展RNA测序（RNA-Seq）表达谱分析。 实验方法：采用Direct-zol RNA MiniPrep试剂盒从快速冷冻的小鼠肝脏中提取并分离总RNA，取100ng RNA作为起始量进行文库制备。文库构建由诺禾致源（Novogene）完成，使用的试剂盒为NEBNext® Ultra RNA Library Prep Kit for Illumina。质检合格的文库在Illumina NovaSeq6000平台上采用双端150bp测序模式完成测序。 实验结果：我们鉴定出不同生殖阶段间胆汁酸生成与FGFR4信号通路存在表达差异，上述两类通路在泌乳期呈现上调。此外，参与促进胆汁酸合成的单个基因在泌乳期表达升高，而与抑制胆汁酸合成相关的基因则在泌乳期表达降低、在泌乳退化阶段表达升高。本数据集的测序样本为7只未产小鼠、6只泌乳期小鼠以及6只泌乳退化第6天小鼠的肝脏来源mRNA，采用Illumina NovaSeq6000平台进行150循环双端测序得到。将得到的Fastq文件比对至mm10小鼠参考基因组（GRCm38.89），基于基因注释文件Mus_musculus.GRCm38.89.chr.gtf，通过RSEM（版本1.3.1）完成单基因计数定量。随后使用DESeq2（版本1.22.2）进行差异基因表达分析。