UFM1类泛素化修饰系统成员E3连接酶UFL1的缺失被证实可引发多种细胞内ER stress，影响细胞增殖或凋亡。卵巢GC增殖、分化和凋亡对卵泡发育起关键作用。我们前期实验发现，敲除UFL1表达造成GC凋亡，闭锁卵泡增多，提示UFL1缺失引发了GC内ER stress，导致卵泡闭锁。本课题拟构建UFL1-/-转基因小鼠，探讨敲除UFL1后造成GC内ER stress，影响卵泡发育的机制。摘取UFL1-/-卵巢，TUNEL、HE，ELISA等检测卵泡内GC，卵泡发育及甾体激素；原代分离GC，qPCR、IB检测ER stress、增殖、分化和凋亡标志分子，CCK8、FCM等检测GC增殖、分化和凋亡；采用UFL1过表达慢病毒、FSH及ER stress阻断剂，观察对GC内ER stress的拯救；采用UFL1-shRNA慢病毒感染WT GC，观察引发ER stress现象。

The E3 ligase UFL1, one of Ubiquitin-like modified system molecular, has been shown to induce multiple intracellular ER stress when it was deleted, which may affect cell proliferation or apoptosis. Ovarian GC proliferation, differentiation and apoptosis play a key role in follicular development. Our pre-experiments showed that Knockout of UFL1 expression leads to apoptosis of GC and increase of atresia follicles, suggesting that UFL1 deletion causes ER stress in GC and leads to follicular atresia. By Construct UFL1-/- knock down mouse with UFL1Loxp/Loxp mouse mating with AMH-Cre, Rosa-Cre-ERT2 under different conditions, our projects intend to explore the mechanism of ER stress in GC and the influence of follicle development absence URFL1. The UFL1-/- GC, follicular development and steroidal hormones will be detected by TUNEL, HE, Elisa; the primary culture UFL1-/- GC will be used to detect the markers of ER stress, proliferation, differentiation and apoptosis by qPCR, IB, and GC proliferation, differentiation and apoptosis will be detected by CCK8,FCM; Lentivirus UFL1, FSH and ER stress inhibitor are used to observe the rescue of ER stress in UFL1-/- GC; and Lentivirus UFL1-shRNA infection is used to induce ER stress in wild type GC.