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.

中文翻译：

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小鼠颗粒细胞中 FSH 调节基因表达和组蛋白修饰的全局分析。

促卵泡激素 (FSH) 通过特定的基因表达程序调节卵巢卵泡发育。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并在用马绒毛膜促性腺激素 (eCG) 刺激之前和 48 小时后收集颗粒细胞。我们使用 RNA 测序（N  = 3/时间点）和使用染色质免疫沉淀和测序（ChIP-Seq；N = 2/时间点）。在整个小鼠基因组中，14,583 个基因具有相关的 H3K4me3 峰，其中 63-66% 的峰出现在≤1 kb 启动子区域内。相对于 0 h eCG，有 72 个基因在 48 h eCG（绝对对数倍数变化 > 1；错误发现率 [FDR] < 0.05）时具有差异的 H3K4me3 修饰。转录组数据分析显示，48 小时心电图有 1463 个差异表达基因（绝对对数倍数变化 > 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具有较高的 H3K4me3 丰度和表达，而Nrip2具有较低的 H3K4me3 丰度和表达。使用 ChIP-qPCR，我们发现 FSH 调节Lhcgr、Cyp19a1、Nppc和Nrip2通过在其各自启动子处调节 H3K4me3。使用 Kallisto-Sleuth 工具进行的转录亚型分析显示 48 小时 eCG 时有 875 个差异表达的转录本（b  > 1；FDR < 0.05）。RNA-seq 数据的通路分析表明，TGF-β 信号传导和类固醇生成通路在 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。这些数据构成了进一步研究调查这些特定机制如何调节颗粒细胞功能的基础。