Abstract
in English, Chinese Zona pellucida glycoprotein-1 (ZP1) is essential for maintaining oocyte structural integrity and facilitating fertilization. Mutations in ZP1 are strongly associated with primary infertility disorders such as fertilization failure and empty follicle syndrome; however, the absence of accurate experimental models has hindered mechanistic understanding and obscured the etiological basis of ZP1-related infertility. In this study, CRISPR/Cas9-mediated genome editing was employed to generate two ZP1-edited cynomolgus macaques ( Macaca fascicularis), designated #ZP1-1 (male) and #ZP1-2 (female). Following sexual maturation, oocytes retrieved from #ZP1-2 through superovulation exhibited a marked increase in zona pellucida-deficient oocytes and a significant reduction in maturation rates compared to controls. Integrated analyses, including immunofluorescence staining, transmission electron microscopy, transcriptomic profiling of oocytes, and histopathological examination of ovarian tissue, revealed disrupted folliculogenesis and oocyte anomalies consistent with phenotypes observed in human empty follicle syndrome. These findings establish the ZP1-knockout cynomolgus macaque as the first non-human primate model of ZP1-related infertility, providing a valuable platform for elucidating disease mechanisms and informing the development of targeted interventions for infertility arising from ZP gene mutations. 透明带糖蛋白-1（ZP1）在保护卵母细胞及受精过程中发挥关键作用。ZP1基因突变与受精障碍、空卵泡综合征等原发性不孕症密切相关，但迄今尚未建立精准模拟ZP1突变致不孕的动物模型，导致其确切病因与机制尚未阐明。该研究利用CRISPR/Cas9技术构建了两只ZP1基因编辑食蟹猴（ Macaca fascicularis），分别命名为#ZP1-1（雄性）和#ZP1-2（雌性）。待#ZP1-2性成熟后，通过超数排卵技术获取其卵母细胞进行评估。研究发现：与对照组相比，该模型卵母细胞透明带缺失比例显著升高，成熟率显著降低。通过对#ZP1-2卵母细胞进行免疫荧光染色、透射电镜观察、转录组测序分析，以及对其卵巢组织进行病理染色，发现其存在原始卵泡发育受损现象，同时表现出与人类空卵泡综合征相似的症状。因此，该研究构建的ZP1基因敲除食蟹猴模型阐明了ZP1缺陷导致不孕的病因，为相关研究提供了关键数据，对ZP基因突变相关不孕症的研究及防治新策略开发具有重要启示意义。.