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The global effect of follicle-stimulating hormone and tumour necrosis factor α on gene expression in cultured bovine ovarian granulosa cells

机译:促卵泡激素和肿瘤坏死因子α对培养的牛卵巢颗粒细胞基因表达的整体影响

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摘要

BackgroundOocytes mature in ovarian follicles surrounded by granulosa cells. During follicle growth, granulosa cells replicate and secrete hormones, particularly steroids close to ovulation. However, most follicles cease growing and undergo atresia or regression instead of ovulating. To investigate the effects of stimulatory (follicle-stimulating hormone; FSH) and inhibitory (tumour necrosis factor alpha; TNFα) factors on the granulosa cell transcriptome, bovine ovaries were obtained from a local abattoir and pools of granulosa cells were cultured in vitro for six days under defined serum-free conditions with treatments present on days 3–6. Initially dose–response experiments (n = 4) were performed to determine the optimal concentrations of FSH (0.33 ng/ml) and TNFα (10 ng/ml) to be used for the microarray experiments. For array experiments cells were cultured under control conditions, with FSH, with TNFα, or with FSH plus TNFα (n = 4 per group) and RNA was harvested for microarray analyses.
机译:背景卵母细胞在颗粒细胞包围的卵泡中成熟。在卵泡生长期间,颗粒细胞复制并分泌激素,尤其是接近排卵的类固醇。然而,大多数卵泡停止生长,并发生闭锁或消退而不是排卵。为了研究刺激性(促卵泡激素; FSH)和抑制性(肿瘤坏死因子α;TNFα)对颗粒细胞转录组的影响,从当地屠宰场获得了牛卵巢,并在体外培养了六个颗粒细胞在规定的无血清条件下的第3天(第3-6天)进行治疗。最初进行剂量-反应实验(n = 4)以确定用于微阵列实验的FSH(0.33 ng / ml)和TNFα(10 ng / ml)的最佳浓度。对于阵列实验,在对照条件下,用FSH,TNFα或FSH加TNFα(每组n = 4)培养细胞,并收集RNA进行微阵列分析。

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