Lei Mingming, Cai Liuping, Li Hui, Chen Zhen, Shi Zhendan
Laboratory of Animal Breeding and Reproduction, Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, China.
Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, China.
Reprod Biol. 2017 Mar;17(1):79-88. doi: 10.1016/j.repbio.2017.01.002. Epub 2017 Jan 9.
Inhibin can regulate granulosa cell proliferation and function via direct action on granulosa cells, or indirectly through stimulation of pituitary follicle-stimulating hormone secretion. Thus far, it has not been possible to unravel or formulate the chain of molecular events that lead to enhanced granulosa cell proliferation and function using conventional gene expression analysis. The aim of this study was to examine the biological effects of immuno-neutralization of inhibin bioactivity in porcine granulosa cells using transcriptome profiling by the RNA-seq technology. Treatment of granulosa cells with anti-inhibin α subunit antibodies increased both cell proliferation and estradiol secretion. Data revealed by RNA sequencing were subjected to bioinformatic analysis. The results showed that a total of 476 genes, including 27 novel genes, were differentially expressed in anti- inhibin antibody-treated granulosa cells compared to untreated granulosa cells. RNA sequencing data were validated by qRT-PCR which confirmed differential expression (upregulation and downregulation) of eighteen of twenty selected genes A total of 476 differentially expressed genes were enriched in processes such as matrix remodeling, chemokine activity, protein binding, and structural molecular activities, and which could be related to granulosa cell proliferation, estradiol synthesis, and ovarian follicle growth. In particular, the data emphasized the importance of extracellular matrix remodeling and the involvement of chemokines in enhanced granulosa cell function, which are important features of ovarian follicle growth, development, maturation, and ovulation. This study provided a new level of understanding of enhanced granulosa cell function and ovarian follicle development achieved through immuno-neutralization of endogenous inhibin bioactivity.
抑制素可通过直接作用于颗粒细胞，或间接通过刺激垂体促卵泡激素分泌来调节颗粒细胞的增殖和功能。迄今为止，使用传统的基因表达分析方法，尚无法阐明或构建导致颗粒细胞增殖和功能增强的分子事件链。本研究的目的是利用RNA测序技术进行转录组分析，研究免疫中和猪颗粒细胞中抑制素生物活性的生物学效应。用抗抑制素α亚基抗体处理颗粒细胞可增加细胞增殖和雌二醇分泌。对RNA测序揭示的数据进行生物信息学分析。结果表明，与未处理的颗粒细胞相比，在抗抑制素抗体处理的颗粒细胞中共有476个基因差异表达，其中包括27个新基因。通过qRT-PCR对RNA测序数据进行验证，证实了20个选定基因中的18个基因的差异表达（上调和下调）。共有476个差异表达基因富集于基质重塑、趋化因子活性、蛋白质结合和结构分子活性等过程中，这些过程可能与颗粒细胞增殖、雌二醇合成和卵泡生长有关。特别是，数据强调了细胞外基质重塑的重要性以及趋化因子在增强颗粒细胞功能中的作用，这些都是卵泡生长、发育、成熟和排卵的重要特征。本研究为通过免疫中和内源性抑制素生物活性实现增强颗粒细胞功能和卵泡发育提供了新的认识水平。
