Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease, Ministry of Agriculture, Hohhot, China.
Theriogenology. 2013 Aug;80(3):248-55. doi: 10.1016/j.theriogenology.2013.04.002. Epub 2013 Jun 5.
The factor associated suicide (Fas) and its ligand (FasL) signaling is an important regulatory pathway of apoptosis in mammalian follicles. However, whether apoptosis in bovine oocytes is regulated by the Fas-FasL signaling pathway remains unknown. In this study, localization of Fas and FasL in immature oocytes and FasL in cumulus cells were examined using immunofluorescence staining. In addition, exogenous FasL was added to an in vitro culture system to investigate apoptotic changes in bovine oocytes, using annexin-V and terminal uridine nick-end labeling staining, and real-time quantitative polymerase chain reaction. In this study, Fas was expressed in immature oocytes, whereas FasL was expressed in cumulus cells, but not in immature oocytes; annexin-V- and terminal uridine nick-end labeling-positive rates of oocytes treated with 2, 10, or 50 ng/mL FasL were higher than those of control oocytes (P < 0.05); and oocytes from the three treatment groups had higher expression levels of Fas and B cell lymphoma/leukemia-2 associated X than those in the control group (P < 0.05). Taken together, we concluded that the Fas-FasL signaling pathway was involved in regulation of bovine oocyte apoptosis, perhaps related to B cell lymphoma/leukemia-2 associated X upregulation.
Fas 及其配体(FasL)信号转导与哺乳动物卵泡细胞凋亡的重要调控途径有关。然而,牛卵母细胞凋亡是否受 Fas-FasL 信号通路调控尚不清楚。本研究采用免疫荧光染色法检测了 Fas 和 FasL 在未成熟卵母细胞中的定位以及 FasL 在卵丘细胞中的定位。此外,还通过 Annexin-V 和末端尿嘧啶核苷酸末端标记染色和实时定量聚合酶链反应,在外源 FasL 添加到体外培养系统中,研究了牛卵母细胞的凋亡变化。本研究中,Fas 在未成熟卵母细胞中表达,而 FasL 在卵丘细胞中表达,但不在未成熟卵母细胞中表达;用 2、10 或 50ng/ml FasL 处理的卵母细胞的 Annexin-V 和末端尿嘧啶核苷酸末端标记阳性率高于对照组卵母细胞(P<0.05);且这三个处理组的卵母细胞中 Fas 和 B 细胞淋巴瘤/白血病-2 相关 X 的表达水平高于对照组(P<0.05)。综上所述,我们得出结论,Fas-FasL 信号通路参与了牛卵母细胞凋亡的调控,可能与 B 细胞淋巴瘤/白血病-2 相关 X 的上调有关。