Reproductive Medical Center, the First Affiliated Hospital of Sun Yat-sen University, No 58, Zhongshan Rd 2, Guangzhou, China.
J Assist Reprod Genet. 2011 Jun;28(6):559-66. doi: 10.1007/s10815-011-9590-0. Epub 2011 Jun 7.
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.
CONCLUSION(S): 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.
鉴定人卵母细胞减数分裂过程中差异表达的 microRNAs（miRNAs）和特定 miRNAs 的表达模式。
通过 miRNA 微阵列分析 GV 期卵母细胞和在常规 FSH 水平（5.5ng/ml）下成熟的 MII 期卵母细胞中差异表达的 miRNAs。实时 RT-PCR 用于验证变化的 miRNAs。为了验证 miRNAs 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 GV 期卵母细胞、MI 期卵母细胞、在常规 FSH 水平下成熟的 MII 期卵母细胞和在高 FSH 水平（2000ng/ml）下成熟的 MII 期卵母细胞。
与 GV 期卵母细胞相比，MII 期卵母细胞中 4 个 miRNAs（hsa-miR-193a-5p、hsa-miR-297、hsa-miR-625 和 hsa-miR-602）上调，11 个 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 和 hsa-miR-15a）下调。对 hsa-miR-15a 和 hsa-miR-20a 表达的 RT-PCR 分析显示，从第 1 组到第 4 组卵母细胞中存在一致的动态变化。
人卵母细胞中的特定 miRNAs 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 的表达出现逆转效应。
