Department of Animal Science, McGill University, Sainte-Anne-de-Bellevue, Qubec, Canada.
Present address: Laboratory of Neuroepigenetics, Department of Health Sciences and Technology of the Swiss Federal Institute of Technology, Medical Faculty of the University of Zürich, Statistical Bioinformatics Group, Swiss Institute of Bioinformatics, Zürich, CH-8057, Switzerland.
Mol Reprod Dev. 2020 Oct;87(10):1082-1096. doi: 10.1002/mrd.23419. Epub 2020 Sep 6.
Follicle-stimulating hormone (FSH) regulates ovarian follicular development through a specific gene expression program. We analyzed FSH-regulated transcriptome and histone modification in granulosa cells during follicular development. We used super-stimulated immature mice and collected granulosa cells before and 48 h after stimulation with equine chorionic gonadotropin (eCG). We profiled the transcriptome using RNA-sequencing (N = 3/time-point) and genome-wide trimethylation of lysine 4 of histone H3 (H3K4me3; an active transcription marker) using chromatin immunoprecipitation and sequencing (ChIP-Seq; N = 2/time-point). Across the mouse genome, 14,583 genes had an associated H3K4me3 peak and 63-66% of these peaks were observed within ≤1 kb promoter region. There were 72 genes with differential H3K4me3 modification at 48 h eCG (absolute log fold change > 1; false discovery rate [FDR] < 0.05) relative to 0 h eCG. Transcriptome data analysis showed 1463 differentially expressed genes at 48 h eCG (absolute log fold change > 1; FDR < 0.05). Among the 20 genes with differential expression and altered H3K4me3 modification, Lhcgr had higher H3K4me3 abundance and expression, while Nrip2 had lower H3K4me3 abundance and expression. Using ChIP-qPCR, we showed that FSH-regulated expression of Lhcgr, Cyp19a1, Nppc, and Nrip2 through regulation of H3K4me3 at their respective promoters. Transcript isoform analysis using Kallisto-Sleuth tool revealed 875 differentially expressed transcripts at 48 h eCG (b > 1; FDR < 0.05). Pathway analysis of RNA-seq data demonstrated that TGF-β signaling and steroidogenic pathways were regulated at 48 h eCG. Thus, FSH regulates gene expression in granulosa cells through multiple mechanisms namely altered H3K4me3 modification and inducing specific transcripts. These data form the basis for further studies investigating how these specific mechanisms regulate granulosa cell functions.
卵泡刺激素 (FSH) 通过特定的基因表达程序调节卵泡的发育。我们分析了卵泡发育过程中颗粒细胞中 FSH 调节的转录组和组蛋白修饰。我们使用超刺激的未成熟小鼠，在使用马绒毛膜促性腺激素 (eCG) 刺激前后 48 小时收集颗粒细胞。我们使用 RNA 测序 (RNA-seq; N = 3/时间点) 和染色质免疫沉淀和测序 (ChIP-Seq; N = 2/时间点) 对转录组进行了分析。在整个小鼠基因组中，有 14583 个基因与 H3K4me3 峰相关，其中 63-66%的峰位于 ≤1kb 启动子区域内。在 48 小时 eCG 时，有 72 个基因的 H3K4me3 修饰发生差异（绝对对数倍变化>1；错误发现率 [FDR] <0.05），与 0 小时 eCG 相比。转录组数据分析显示，在 48 小时 eCG 时，有 1463 个基因表达差异（绝对对数倍变化>1；FDR <0.05）。在具有差异表达和改变的 H3K4me3 修饰的 20 个基因中，Lhcgr 的 H3K4me3 丰度和表达更高，而 Nrip2 的 H3K4me3 丰度和表达更低。使用 ChIP-qPCR，我们表明 FSH 通过调节其各自启动子上的 H3K4me3 来调节 Lhcgr、Cyp19a1、Nppc 和 Nrip2 的表达。使用 Kallisto-Sleuth 工具进行的转录本异构体分析显示，在 48 小时 eCG 时，有 875 个转录本表达差异（b>1；FDR <0.05）。RNA-seq 数据的通路分析表明，TGF-β 信号通路和类固醇生成途径在 48 小时 eCG 时受到调节。因此，FSH 通过多种机制调节颗粒细胞中的基因表达，即改变 H3K4me3 修饰和诱导特定转录本。这些数据为进一步研究这些特定机制如何调节颗粒细胞功能奠定了基础。
