New insights into the pathogenesis of cystic follicles in cattle: Microarray analysis of gene expression in granulosa cells
New insights into the pathogenesis of cystic follicles in cattle: Microarray analysis of gene expression in granulosa cells
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DOI：
10.2527/jas.2010-3463
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发表时间：
2011-06-01
影响因子：
3.3
通讯作者：
Spicer, L. J.
中科院分区：
文献类型：
--
作者：
Grado-Ahuir, J. A.;Aad, P. Y.;Spicer, L. J.
卵巢卵泡的生长发育受卵巢内外因素的调控，影响颗粒细胞的增殖和分化。然而，驱动卵泡生长的分子机制尚未完全了解。卵巢滤泡囊肿是导致奶牛繁殖失败的最常见原因之一。然而，囊肿形成的主要原因尚未明确。基因表达比较可能有助于阐明卵巢囊肿疾病的原因。我们的目的是确定差异表达的基因在卵巢颗粒细胞之间的正常显性和囊性卵泡的牛。通过超声引导抽吸从奶牛（n = 24）或肉牛（n = 23）的屠宰场卵巢收集的优势和囊性卵泡中分离颗粒细胞和卵泡液。用放射免疫分析法测定卵泡液中孕酮、雌二醇和雄烯二酮。提取总RNA，并与6个Affyssin GeneChip牛基因组阵列（Affyssin，Santa Clara，CA）杂交。通过实时定量逆转录PCR检测差异表达基因的mRNA表达。与非囊性卵泡相比，囊性卵泡中孕酮含量较高（P <0.05），雌二醇含量较低（P < 0.05），雄烯二酮含量无差异（P > 0.10）。共发现163条差异表达基因序列（P < 0.01），其中19条上调，144条下调。从选定的靶基因，定量实时逆转录PCR证实血管生成素，PGE（2）受体4，和G蛋白偶联受体34基因在囊性卵泡中上调，和印度刺猬蛋白前体和分泌卷曲相关蛋白4基因在囊性卵泡中下调。需要进一步的研究来阐明这些因子在卵泡发育和囊肿形成中的作用。
Ovarian follicular growth and development are regulated by extraovarian and intraovarian factors, which influence granulosa cell proliferation and differentiation. However, the molecular mechanisms that drive follicular growth are not completely understood. Ovarian follicular cysts are one of the most common causes of reproductive failure in dairy cattle. Nevertheless, the primary cause of cyst formation has not been clearly established. A gene expression comparison may aid in elucidating the causes of ovarian cyst disease. Our objective was to identify differentially expressed genes in ovarian granulosa cells between normal dominant and cystic follicles of cattle. Granulosa cells and follicular fluid were isolated from dominant and cystic follicles collected via either ultrasound-guided aspiration from dairy cows (n = 24) or slaughterhouse ovaries from beef cows (n = 23). Hormonal analysis for progesterone, estradiol, and androstenedione in follicular fluid was performed by RIA. Total RNA was extracted and hybridized to 6 Affymetrix GeneChip Bovine Genome Arrays (Affymetrix, Santa Clara, CA). Abundance of mRNA for differentially expressed selected genes was determined through quantitative realtime reverse-transcription PCR. Follicular cysts showed greater (P < 0.05) progesterone, lesser (P < 0.05) estradiol, and no differences (P > 0.10) in androstenedione concentrations compared with noncystic follicles. A total of 163 gene sequences were differentially expressed (P < 0.01), with 19 upregulated and 144 downregulated. From selected target genes, quantitative real-time reverse-transcription PCR confirmed angiogenin, PGE(2) receptor 4, and G-protein coupled receptor 34 genes as upregulated in cystic follicles, and Indian hedgehog protein precursor and secreted frizzled-related protein 4 genes as downregulated in cystic follicles. Further research is required to elucidate the role of these factors in follicular development and cyst formation.