Abstract：
The objective of this study was to examine the expression patterns of secreted frizzled-related protein 5 (SFRP5) in the ovaries of Hu sheep and to explore the key downstream factors of SFRP5 in sheep granulosa cells (GCs) using RNA-seq. In the present study, SFRP5 was widely expressed in the ovary and localized to GCs and oocytes during various stages of follicular development. In addition, the expression of SFRP5 increased with follicular diameter. In contrast to the negative control, SFRP5 knockdown promoted the EdU-positive cell rate with an increase in PCNA mRNA and protein levels, whereas SFRP5 overexpression had the opposite effect. In addition, the cell cycle was propelled from the G0/G1 phase to the S phase with the upregulation of CCNB1, CCND1, CDK1, and CDK4 after SFRP5 knockdown. Moreover, SFRP5 overexpression enhanced the apoptosis of GCs with increased Caspase3 protein levels. Following SFRP5 knockdown, differentially expressed genes were mainly enriched in the PI3K/AKT, MAPK, Wnt, and Hippo signaling pathways, and several related candidate genes such as MMP1, MMP3, SFRP4, INHA, TGFA, and CASP3 were screened. In general, this study enhances our understanding of the expression of SFRP5 in the GCs of Hu sheep, along with its functions in follicular development.
摘要：
本研究的目的是检测分泌型卷曲相关蛋白5（SFRP5）在湖羊卵巢中的表达模式，并利用RNA-seq探索SFRP5在绵羊颗粒细胞（GCs）中的关键下游因子。在本研究中,SFRP5在卵巢中广泛表达，并在卵泡发育的各个阶段定位于GC和卵母细胞。此外,SFRP5的表达随卵泡直径的增加而增加。与阴性对照相比，SFRP5敲低可促进EdU阳性细胞率，增加PCNAmRNA和蛋白水平，而SFRP5过表达具有相反的作用。此外,随着SFRP5敲低后CCNB1,CCND1,CDK1和CDK4的上调,细胞周期从G0/G1期被推进至S期.此外，SFRP5过表达通过增加Caspase3蛋白水平增强GCs的凋亡。SFRP5击倒后，差异表达基因主要富集在PI3K/AKT,MAPK,Wnt,和河马信号通路，和几个相关的候选基因，如MMP1，MMP3，SFRP4，INHA，TGFA,对CASP3进行筛选。总的来说,这项研究增强了我们对SFRP5在湖羊GCs中表达的理解，及其在卵泡发育中的功能。
