Differentially expressed micoRNAs in human oocytes
Differentially expressed micoRNAs in human oocytes
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DOI：
10.1007/s10815-011-9590-0
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发表时间：
2011-06-01
影响因子：
3.1
通讯作者：
Zhou, Canquan
中科院分区：
文献类型：
--
作者：
Xu, Yan-Wen;Wang, Bin;Zhou, Canquan
为了鉴定人卵母细胞减数分裂过程中差异表达的 microRNA (miRNA) 和特定 miRNA 的表达模式。为了鉴定差异表达的 miRNA，通过 miRNA 微阵列分析在常规 FSH 水平 (5.5 ng/ml) 下成熟的 GV 卵母细胞和 MII 卵母细胞。使用实时RT-PCR来确认改变的miRNA。为了验证 miRNA 从 GV 到 MII 阶段的动态变化，将卵母细胞分为四组 (#1-4)，分别对应于 GV 卵母细胞、MI 卵母细胞、常规 FSH 水平成熟的 MII 卵母细胞和高 FSH 水平 (2,000 ng/ml) 成熟的 MII 卵母细胞。与 GV 卵母细胞相比，MII 卵母细胞表现出 4 个 miRNA (hsa-miR-193a-5p、 hsa-miR-297、hsa-miR-625 和 hsa-miR-602），以及 11 个 miRNA（hsa-miR-888*、hsa-miR-212、hsa-miR-662、hsa-miR-299-5p、hsa-miR-339-5p、hsa-miR-20a、hsa-miR-486-5p、 hsa-miR-141*、hsa-miR-768-5p、hsa-miR-376a 和 hsa-miR-15a)。 hsa-miR-15a 和 hsa-miR-20a 表达的 RT-PCR 分析显示，第 1 组到第 4 组卵母细胞的动态变化一致。人类卵母细胞中的特定 miRNA 在减数分裂过程中发生动态变化。 IVM 培养基中高浓度 FSH 导致 hsa-miR-15a 和 hsa-miR-20a 表达逆转。
To identify differentially expressed microRNAs (miRNAs) and expression patterns of specific miRNAs during meiosis in human oocytes.To identify differentially expressed miRNAs, GV oocytes and MII oocytes matured at conventional FSH levels (5.5 ng/ml) were analyzed by miRNA microarray. Real-time RT-PCR was used to confirm the changed miRNAs. To validate the dynamic changes of miRNAs from GV to MII stages, oocytes were divided into four groups (#1-4), corresponding to GV oocytes, MI oocytes, MII oocytes matured in conventional FSH level and MII oocytes matured in high FSH level (2,000 ng/ml) respectively.Compared with GV oocytes, MII oocytes exhibited up-regulation of 4 miRNAs (hsa-miR-193a-5p, hsa-miR-297, hsa-miR-625 and hsa-miR-602), and down-regulation of 11 miRNAs (hsa-miR-888*, hsa-miR-212, hsa-miR-662, hsa-miR-299-5p, hsa-miR-339-5p, hsa-miR-20a, hsa-miR-486-5p, hsa-miR-141*, hsa-miR-768-5p, hsa-miR-376a and hsa-miR-15a). RT-PCR analysis of hsa-miR-15a and hsa-miR-20a expression revealed concordant dynamic changes in oocytes from group 1 to group 4.Specific miRNAs in human oocytes had dynamic changes during meiosis. High-concentration FSH in IVM medium led to reverse effect on the expression of hsa-miR-15a and hsa-miR-20a.