Schütz L F, Hurst R E, Schreiber N B, Spicer L J
Department of Animal Science, Oklahoma State University, Stillwater, OK 74078, USA.
Department of Urology, University of Oklahoma College of Medicine, Oklahoma City, OK 73104, USA.
Domest Anim Endocrinol. 2018 Apr;63:48-58. doi: 10.1016/j.domaniend.2017.12.002. Epub 2018 Jan 4.
We reported previously that fibroblast growth factor 9 (FGF9) acts as an antidifferentiation factor, stimulating proliferation of granulosa cells (GCs) and theca cells (TCs) while suppressing hormone-induced steroidogenesis of these cells. How FGF9 acts to simultaneously suppress steroidogenesis and stimulate proliferation remains to be fully elucidated. Thus, this study was undertaken to clarify the effects of FGF9 on the TC transcriptome. Ovaries were obtained from beef heifers at a local abattoir, TCs were isolated from large antral follicles, and cultured with or without 30 ng/mL of FGF9 for 24 h in the presence of LH and IGF-1. After treatment, total RNA was extracted from TC and processed for microarray using Affymetrix GeneChip Bovine Genome Arrays (n = 4/group). Transcriptome analysis comparing FGF9-treated TC with control TC using 1.3-fold cutoff, and a P < 0.05 significance level identified 355 differentially expressed transcripts, with 164 elements upregulated and 191 elements downregulated by FGF9. The ingenuity pathway analysis (IPA) was used to investigate how FGF9 treatment affects molecular pathways, biological functions, and the connection between molecules in bovine TC. The IPA software identified 346 pathways in response to FGF9 in TC involved in several biological functions and unveiled interesting relationships among genes related to cell proliferation (eg, CCND1, FZD5, and MYB), antioxidation/cytoprotection (eg, HMOX1 and NQO1), and steroidogenesis (eg, CYP11A1 and STAR). Overall, genes, pathways, and networks identified in this study painted a picture of how FGF9 may regulate folliculogenesis, providing novel candidate genes for further investigation of FGF9 functions in ovarian follicular development.
我们之前报道过，成纤维细胞生长因子9（FGF9）作为一种抗分化因子，可刺激颗粒细胞（GCs）和卵泡膜细胞（TCs）增殖，同时抑制这些细胞的激素诱导类固醇生成。FGF9如何同时抑制类固醇生成并刺激增殖仍有待充分阐明。因此，本研究旨在阐明FGF9对TC转录组的影响。从当地屠宰场的肉用小母牛获取卵巢，从大的窦状卵泡中分离出TCs，并在促黄体生成素（LH）和胰岛素样生长因子-1（IGF-1）存在的情况下，用或不用30 ng/mL的FGF9培养24小时。处理后，从TC中提取总RNA，并使用Affymetrix GeneChip牛基因组阵列（每组n = 4）进行微阵列分析。转录组分析以1.3倍的截断值和P < 0.05的显著性水平比较FGF9处理的TC与对照TC，鉴定出355个差异表达的转录本，其中164个元件被FGF9上调，191个元件被下调。 Ingenuity通路分析（IPA）用于研究FGF9处理如何影响牛TC中的分子通路、生物学功能以及分子之间的联系。IPA软件在TC中鉴定出346条对FGF9有反应的通路，这些通路涉及多种生物学功能，并揭示了与细胞增殖（如CCND1、FZD5和MYB）、抗氧化/细胞保护（如HMOX1和NQO1）以及类固醇生成（如CYP11A1和STAR）相关的基因之间有趣的关系。总体而言，本研究中鉴定出的基因、通路和网络描绘了FGF9可能如何调节卵泡发生的图景，为进一步研究FGF9在卵巢卵泡发育中的功能提供了新的候选基因。
