Gene expression analysis of pig cumulus-oocyte complexes stimulated in vitro with follicle stimulating hormone or epidermal growth factor-like peptides.
Gene expression analysis of pig cumulus-oocyte complexes stimulated in vitro with follicle stimulating hormone or epidermal growth factor-like peptides.
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DOI：
10.1186/s12958-015-0112-2
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发表时间：
2015-10-06
期刊：
影响因子：
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通讯作者：
Prochazka R
中科院分区：
文献类型：
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作者：
Blaha M;Nemcova L;Kepkova KV;Vodicka P;Prochazka R
促性腺激素诱导的排卵前卵泡卵母细胞减数分裂的恢复是由表皮生长因子（EGF）样肽，双调蛋白（AREG）和epiregulin（EREG），在壁颗粒和卵丘细胞的表达。促性腺激素和EGF样肽都具有通过激活卵丘细胞中广泛的信号网络来刺激体外卵母细胞减数分裂恢复的能力。为了更好地了解促性腺激素（FSH）和EGF样肽的快速基因组作用，我们分析了卵丘细胞刺激后3小时的转录组。我们将卵丘细胞的aRNA与猪寡核苷酸微阵列杂交，并将FSH和AREG/EREG刺激的卵丘细胞的转录组与未处理的对照细胞进行比较，反之亦然。根据其分子和细胞功能，对鉴定的过表达和低表达基因进行功能基因组学分析。通过实时qRT-PCR验证了50个在卵巢发育中具有已知或潜在功能的选定基因的表达模式。FSH和AREG/EREG都增加了与细胞增殖、细胞迁移、血液凝固和细胞外基质重塑调节相关的基因的表达。FSH单独诱导参与炎症反应和活性氧反应的基因表达。此外，FSH刺激与一些排卵事件密切相关的基因的表达，这些基因的表达完全或显著高于AREG/EREG（AREG、ADAMTS 1、HAS 2、TNFAIP 6、PLAUR、PLAT和HSD 17 B7）。与AREG/EREG相反，FSH还增加了编码关键转录因子（CEBPB，FOS，ID 1/3和NR 5A 2）的基因的表达，这可能有助于FSH和AREG/EREG处理的卵丘细胞的不同表达谱。FSH对卵丘细胞基因转录的影响在受影响的细胞功能数量以及过表达和低表达基因数量方面高于EGF样因子的影响。FSH和EGF样因子均过表达参与类固醇生成和组织重塑中排卵后转换的基因。然而，FSH在上调成熟卵母细胞排卵所必需的几个特定基因以及在卵丘封闭卵母细胞体外成熟中起重要作用的基因方面显着更有效。本文的在线版本（doi：10.1186/s12958-015-0112-2）包含补充材料，可供授权用户使用。
The gonadotropin-induced resumption of oocyte meiosis in preovulatory follicles is preceded by expression of epidermal growth factor (EGF)-like peptides, amphiregulin (AREG) and epiregulin (EREG), in mural granulosa and cumulus cells. Both the gonadotropins and the EGF-like peptides possess the capacity to stimulate resumption of oocyte meiosis in vitro via activation of a broad signaling network in cumulus cells. To better understand the rapid genomic actions of gonadotropins (FSH) and EGF-like peptides, we analyzed transcriptomes of cumulus cells at 3 h after their stimulation. We hybridized aRNA from cumulus cells to a pig oligonucleotide microarray and compared the transcriptomes of FSH- and AREG/EREG-stimulated cumulus cells with untreated control cells and vice versa. The identified over- and underexpressed genes were subjected to functional genomic analysis according to their molecular and cellular functions. The expression pattern of 50 selected genes with a known or potential function in ovarian development was verified by real-time qRT-PCR. Both FSH and AREG/EREG increased the expression of genes associated with regulation of cell proliferation, cell migration, blood coagulation and extracellular matrix remodeling. FSH alone induced the expression of genes involved in inflammatory response and in the response to reactive oxygen species. Moreover, FSH stimulated the expression of genes closely related to some ovulatory events either exclusively or significantly more than AREG/EREG (AREG, ADAMTS1, HAS2, TNFAIP6, PLAUR, PLAT, and HSD17B7). In contrast to AREG/EREG, FSH also increased the expression of genes coding for key transcription factors (CEBPB, FOS, ID1/3, and NR5A2), which may contribute to the differing expression profiles of FSH- and AREG/EREG-treated cumulus cells. The impact of FSH on cumulus cell gene transcription was higher than the impact of EGF-like factors in terms of the number of cell functions affected as well as the number of over- and underexpressed genes. Both FSH and EGF-like factors overexpressed genes involved in the post-ovulatory switch in steroidogenesis and tissue remodelling. However, FSH was remarkably more efficient in the up-regulation of several specific genes essential for ovulation of matured oocytes and also genes that been reported to play an important role in maturation of cumulus-enclosed oocytes in vitro. The online version of this article (doi:10.1186/s12958-015-0112-2) contains supplementary material, which is available to authorized users.