Huang Lin, Hou Yuanyuan, Li Hu, Wu Hanxiao, Hu Jianing, Lu Yangqing, Liu Xingting
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Animal Science and Technology, Guangxi University, 530004, Nanning, China.
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Animal Science and Technology, Guangxi University, 530004, Nanning, China.
Theriogenology. 2022 May;184:140-152. doi: 10.1016/j.theriogenology.2022.03.012. Epub 2022 Mar 15.
During the hierarchical development of follicles in laying hens, a large number of small white follicles (SWFs) undergo atresia caused by apoptosis rather than growing into large white follicles (LWFs), which reduces the number of follicles developing into hierarchical preovulatory follicles and results in declines in egg production. Therefore, figuring out factors responsible for SWFs atresia is necessary to improve egg production of laying hens. Endoplasmic reticulum stress (ERS) is one of the possible causes of cell apoptosis. However, the association between SWFs atresia and ERS in hens is yet to be answered. In this study, cell proliferation and apoptosis, expression levels of proteins and genes related to ERS, and cell ultrastructure were compared to elucidate whether ERS occurred in atretic small white follicles (ASWFs). Subsequently, a tunicamycin-induced ERS model and a serum withdrawal-induced atresia model of SWFs in vitro were established to investigate whether ERS could lead to follicular atresia. The results showed that the cell apoptosis increased significantly while the cell proliferation decreased evidently in ASWFs compared with SWFs. Meanwhile, swollen and vacuolization deformation of ERs was observed in the GCs but not TCs of ASWFs. Results of experiments in vitro demonstrated that treatment with serum withdrawal or tunicamycin inhibited cell proliferation and promoted cell apoptosis in SWFs, and the adverse effects of tunicamycin on SWFs manifested a dose-dependent increase. In comparison with the controls, treatment with serum withdrawal or tunicamycin upregulated expression levels of proteins and genes in ERS pathway. Moreover, the dose of 5 μg/mL tunicamycin seemed to be the optimal concentration for establishing the tunicamycin-induced ERS model. Meanwhile, the results of cell ultrastructure observation showed that ERs in GCs of SWFs of serum withdrawal group and treatment with 5 μg/mL tunicamycin group became swollen, dilatation and vacuolization. In conclusion, the data of this study indicated that ERS is involved in SWF atresia in laying hens, and ERS in GCs might be more severe than that in TCs.
在蛋鸡卵泡的分级发育过程中,大量小白卵泡(SWF)因凋亡而发生闭锁,而非发育成大白卵泡(LWF),这减少了发育成分级排卵前卵泡的卵泡数量,导致产蛋量下降。因此,找出导致SWF闭锁的因素对于提高蛋鸡产蛋量至关重要。内质网应激(ERS)是细胞凋亡的可能原因之一。然而,母鸡SWF闭锁与ERS之间的关联尚待明确。在本研究中,比较了细胞增殖和凋亡、与ERS相关的蛋白质和基因的表达水平以及细胞超微结构,以阐明闭锁小白卵泡(ASWF)中是否发生了ERS。随后,建立了衣霉素诱导的ERS模型和体外血清饥饿诱导的SWF闭锁模型,以研究ERS是否会导致卵泡闭锁。结果显示,与SWF相比,ASWF中的细胞凋亡显著增加,而细胞增殖明显减少。同时,在ASWF的颗粒细胞(GC)中观察到内质网肿胀和空泡化变形,但在膜细胞(TC)中未观察到。体外实验结果表明,血清饥饿或衣霉素处理可抑制SWF的细胞增殖并促进细胞凋亡,衣霉素对SWF的不利影响呈剂量依赖性增加。与对照组相比,血清饥饿或衣霉素处理上调了ERS途径中蛋白质和基因的表达水平。此外,5μg/mL衣霉素似乎是建立衣霉素诱导的ERS模型的最佳浓度。同时,细胞超微结构观察结果显示,血清饥饿组和5μg/mL衣霉素处理组的SWF的GC中的内质网变得肿胀、扩张和空泡化。总之,本研究数据表明ERS参与了蛋鸡SWF闭锁,且GC中的ERS可能比TC中的更严重。