Ovarian Stroma RNA-seq of Progesterone receptor null vs WT mice at 8hrs post hCG

Progesterone receptor-dependent expression of the ovairan stroma is investigated to determie PGR control of stromal functions at ovulation. Overall design: Ovaries were collected from 21-25day old PGR(-/-) and PGR(+/+) littermates at 8hrs post-hCG. N=3-4 per genotype with each experimental replicate consisting of tissue pooled from 3 mice. The Stromal tissue of the ovary (GC-depleted ovary) was used for mRNA-seq Illumina Truuseg Strandard mRNA kit, Illumina Novaseq 600 SP. Paired end, 100bp, depth of 54-72M reads. Qualisty control checked using FastQC. Salmon lignment to GRCm39 M26 mouse transcriptome. DESeq2 used to asses differentially expressed genes.

本研究旨在探究孕酮受体（Progesterone Receptor, PGR）依赖的卵巢基质表达模式，以明确PGR对排卵过程中基质细胞功能的调控作用。整体实验设计：于绒毛膜促性腺激素（human chorionic gonadotropin, hCG）处理后8小时，从21至25日龄的PGR敲除（PGR(-/-)）与野生型（PGR(+/+)）同窝仔鼠中采集卵巢组织；每种基因型设置3~4个生物学重复，每个实验重复的组织来源于3只小鼠的混合组织。提取卵巢基质组织（即去除颗粒细胞（granulosa cells, GC）的卵巢组织），使用Illumina TruSeq Standard mRNA建库试剂盒进行mRNA测序，测序平台为Illumina NovaSeq 600 SP；测序模式为双端测序，读长100bp，测序数据量为54~72百万条reads。使用FastQC进行测序数据质量控制，通过Salmon将测序reads比对至GRCm39 M26版本的小鼠转录组，并采用DESeq2分析差异表达基因。