内质网稳态关键因子WFS1在猪卵母细胞成熟与植入前胚胎发育中的作用及其机制研究

内质网是真核细胞中蛋白质合成及钙储存的关键细胞器，在保证细胞内环境稳定、促进细胞增殖与分化中起着重要作用。内质网稳态关键因子WFS1在调节内质网内环境及减少内质网应激中发挥关键作用。然而，WFS1能否通过调控内质网稳态进而影响卵母细胞成熟及植入前胚胎发育尚不清楚。本项目在阐明WFS1在猪卵母细胞及植入前胚胎中的表达规律的基础上，通过siRNA干扰WFS1的表达，检测卵母细胞减数分裂进程、植入前胚胎发育能力及合子基因组激活、胞内钙离子水平、内质网应激、氧化还原能力、DNA损伤修复和细胞凋亡相关标志物的动态变化情况，意在解析WFS1在猪卵母细胞成熟及植入前胚胎发育中的相关通路，阐明其调控机制。研究结果将有助于我们深入了解卵母细胞成熟及植入前胚胎发育过程中的重要事件和调控机制，为明确卵母细胞成熟及植入前胚胎发育阻滞的原因提供理论基础，并为优化卵母细胞及植入前胚胎体外培养体系提供理论支持。

Endoplasmic reticulum (ER) is an important organelle in protein synthesis and calcium storage in eukaryotic cells, and plays a vital role in stabilizing intracellular environment and promoting cell proliferation and differentiation. ER homeostasis key factor WFS1 plays an important role in regulating environment of ER and reducing ER stress. However, it is not clear whether WFS1 could affect oocyte maturation and pre-implantation embryo development by regulating ER homeostasis. Hence, this project aims to clarify the expression pattern of WFS1 in porcine oocyte and pre-implantation embryo, and to analyze the relevant pathways of WFS1 during porcine oocyte maturation and pre-implantation embryo development and clarify its regulation mechanism through siRNA interference with WFS1 expression, which includes detection of oocyte meiosis process and pre-implantation embryo development competence, and evaluate dynamic change of zygotic genome activation, intracellular calcium levels, ER stress, oxidative stress, repair of DNA damage and cell apoptosis markers. These results will help us understand the important events and regulatory mechanisms during oocyte maturation and pre-implantation development. It will not only provide a theoretical basis for finding out the reason that causes the blocking of oocyte maturation and pre-implantation embryo development, but will also provide theoretical support to optimize oocyte and pre-implantation embryo culture system.