Centre de Recherche en Biologie de la Reproduction, Dèpartement des Sciences Animales, Université Laval, Quebec, Quebec, Canada.
Theriogenology. 2012 Jun;77(9):1928-38. doi: 10.1016/j.theriogenology.2012.01.015. Epub 2012 Mar 22.
The objective was to analyze the impact of follicle stimulating hormone (FSH, ovarian stimulation) on the transcriptome of in vivo bovine oocytes three times around the luteinizing hormone (LH) surge. In vivo bovine oocytes were collected 2 h pre-LH surge, 6 h post-LH surge, and 22 h post-LH surge in both naturally ovulating and superovulated animals. To assess potential changes in gene levels, samples were hybridized using a custom bovine microarray. Two series of hybridizations were performed: the first comparing natural vs. stimulated cycles, the second according to time of collection. Among the potential candidates, 13 genes were selected according to their degree of differential expression and their potential link to oocyte competence. Measurements of their relative mRNA levels was made using QPCR. Gene candidates BTG4 (P = 0.0006), PTTG1 (P = 0.0027), PAPOLA (P = 0.0245), and LEO1 (P = 0.0393) had higher mRNA levels in oocytes treated with FSH for all collection times when compared to oocytes produced through the natural cycle. Among our selected candidates, only one gene, GDF9 (P = 0.0261), was present at a higher level in oocytes collected at -2 h and 6 h than 22 h post-LH for all treatments, regardless of the presence of FSH. Although the number of genes influenced by ovarian stimulation seemed low, the observed differences occurred at a time of minimal transcriptional activity and supported the potential impact on the future embryo. These impacts could have been epigenetic in nature, as embryo quality was not reported to be different from stimulated animals.
目的是分析卵泡刺激素 (FSH,卵巢刺激) 对黄体生成素 (LH) 峰前后三次活体牛卵母细胞转录组的影响。在自然排卵和超排卵动物中,分别在 LH 峰前 2 小时、LH 峰后 6 小时和 LH 峰后 22 小时采集活体牛卵母细胞。为了评估基因水平的潜在变化,使用定制的牛微阵列对样本进行杂交。进行了两个系列的杂交:第一个比较自然与刺激周期,第二个根据采集时间。在潜在的候选基因中,根据差异表达程度及其与卵母细胞能力的潜在联系,选择了 13 个基因。使用 QPCR 测量其相对 mRNA 水平。BTG4(P = 0.0006)、PTTG1(P = 0.0027)、PAPOLA(P = 0.0245)和 LEO1(P = 0.0393)等候选基因的 mRNA 水平在所有采集时间点的 FSH 处理卵母细胞中均高于自然周期产生的卵母细胞。在我们选择的候选基因中,只有一个基因 GDF9(P = 0.0261)在所有处理中,LH 峰后-2 小时和 6 小时采集的卵母细胞中的表达水平高于 22 小时,无论是否存在 FSH。尽管受卵巢刺激影响的基因数量似乎较少,但观察到的差异发生在转录活性最小的时期,支持对未来胚胎的潜在影响。这些影响可能是表观遗传的,因为胚胎质量与刺激动物没有不同。