Douville Gabriel, Sirard Marc-André
Centre de Recherche en Biologie de la Reproduction, Faculté des Sciences de l'Agriculture et de l'Alimentation, Département des Sciences Animales, Pavillon INAF, Université Laval, Québec G1V 0A6, QC, Canada.
J Ovarian Res. 2014 May 7;7:50. doi: 10.1186/1757-2215-7-50. eCollection 2014.
The objective of this study was to build the transcriptomic profile of granulosa cells originating from follicles 6 to 9 mm in diameter in dairy cattle using microarrays.
Granulosa cells originating from three different phases of antral follicle growth were compared: growing (G), plateau (P) and atresia (A), as categorized by flow cytometry profiles of DNA. The growing and atretic conditions were each hybridized against the plateau condition as a reference in order to understand the specific biological mechanisms modulated in this class of follicles.
2,942 genes were differentially expressed (P < 0.05) in P vs. G and 1,974 in A vs. P. A clear segregation of the 3 phases was confirmed by between group analysis (BGA). The first characteristic of the plateau phase is the activation of the upstream regulators TP53 and PTEN which participate in the reduction of cell growth through MYC, FOS and E2F1-2-3. We also observed the down-regulation of steroidogenesis genes: CYP11A1 and CYP19A1, in the granulosa cells of the plateau phase relative to the growth phase. On the other hand, the A vs. P contrast showed up-regulation of multiple transcripts associated to apoptosis: CCT2, DAB2, DSG2 and TGM2.
This study offers multiple candidate genes to be further studied in order to elucidate their role in the modulation of follicular development and, ultimately, of oocyte quality.
本研究的目的是使用微阵列构建源自直径6至9毫米卵泡的奶牛颗粒细胞的转录组图谱。
比较了源自窦卵泡生长三个不同阶段的颗粒细胞:生长阶段(G)、平台期(P)和闭锁阶段(A),通过DNA的流式细胞术图谱进行分类。将生长阶段和闭锁阶段的样本分别与平台期样本进行杂交,以作为对照,从而了解这类卵泡中调节的特定生物学机制。
在P组与G组之间有2942个基因差异表达(P < 0.05),在A组与P组之间有1974个基因差异表达。通过组间分析(BGA)证实了这三个阶段的明显分离。平台期的第一个特征是上游调节因子TP53和PTEN的激活,它们通过MYC、FOS和E2F1 - 2 - 3参与细胞生长的减少。我们还观察到,相对于生长阶段,平台期颗粒细胞中类固醇生成基因CYP11A1和CYP19A1的表达下调。另一方面,A组与P组的对比显示,与凋亡相关的多个转录本上调:CCT2、DAB2、DSG2和TGM2。
本研究提供了多个候选基因以供进一步研究,以阐明它们在调节卵泡发育以及最终在调节卵母细胞质量中的作用。