Pan Zezheng, Sun Mengli, Li Jia, Zhou Fangyue, Liang Xia, Huang Jian, Zheng Tuochen, Zheng Liping, Zheng Yuehui
Cell Physiol Biochem. 2015;37(6):2311-22. doi: 10.1159/000438586. Epub 2015 Dec 4.
BACKGROUND/AIMS: Ovarian germline stem cells (OGSCs) have been shown to mainly exist in the ovarian surface epithelium (OSE), but the activity changes of germline stem cells during different reproductive stages and the potential regulatory signaling pathway are still unknown. The Notch signaling pathway plays a key role in cell development, primordial follicles and stem cell proliferation. However, whether it plays a role in the proliferation of OGSCs is unknown. Here, we analyzed the activity changes of germline stem cells and the correlation between germline stem cells and the Notch signaling pathway.
The expression of germline stem cell markers Mvh, Ooc4 and the Notch molecules Notch1, Hes1, and Hes5 were detected during 3 days (3d), and 2, 12, 20 months (2m, 12m, 20m) mouse ovarian surface epithelium samples. DAPT, a specific inhibitor of the Notch pathway, was used to observe the influence of Notch signaling in the germline stem cells.
The results showed that the levels of MVH and OCT4 decreased substantially with reproductive age in ovarian surface epithelium, and the same tendency was detected in the Notch signaling molecules Notch1, Hes1 and Hes5. Dual-IF results showed that the germline stem cell markers were co-expressed with Notch molecules in the ovarian surface epithelium. While, the expression of MVH and OCT4 were reduced when the ovaries were treated with DAPT and the levels were attenuated with increasing dose of DAPT.
Taken together, our results indicate that the viability of OGSCs decreased with the age of the mouse ovaries, and the activity of OGSCs in the ovarian surface epithelium may be related to the Notch signaling pathway.
背景/目的:卵巢生殖系干细胞(OGSCs)主要存在于卵巢表面上皮(OSE)中,但不同生殖阶段生殖系干细胞的活性变化及其潜在的调控信号通路仍不清楚。Notch信号通路在细胞发育、原始卵泡和干细胞增殖中起关键作用。然而,它是否在OGSCs的增殖中发挥作用尚不清楚。在此,我们分析了生殖系干细胞的活性变化以及生殖系干细胞与Notch信号通路之间的相关性。
在3天(3d)、2、12、20个月(2m、12m、20m)的小鼠卵巢表面上皮样本中检测生殖系干细胞标志物Mvh、Ooc4以及Notch分子Notch1、Hes1和Hes5的表达。使用Notch通路的特异性抑制剂DAPT来观察Notch信号对生殖系干细胞的影响。
结果显示,卵巢表面上皮中MVH和OCT4的水平随着生殖年龄的增加而显著降低,Notch信号分子Notch1、Hes1和Hes5也呈现相同的趋势。双免疫荧光结果显示,生殖系干细胞标志物与卵巢表面上皮中的Notch分子共表达。同时,用DAPT处理卵巢后,MVH和OCT4的表达降低,且随着DAPT剂量的增加而减弱。
综上所述,我们的结果表明,OGSCs的活力随着小鼠卵巢年龄的增长而降低,卵巢表面上皮中OGSCs的活性可能与Notch信号通路有关。