Abstract：
Because of the time-consuming nature of surgical neutering and the rapid rate of reproduction among domestic cats, it is crucial to investigate alternative, nonsurgical methods of contraception for this species. Sperm protein IZUMO1 and its oocyte receptor JUNO have been proposed as potential targets for nonsurgical contraceptives. This study aimed to demonstrate (1) the protein coding sequence of feline IZUMO1 and JUNO, (2) gene expression in specific organs by measuring mRNA levels in different visceral tissues, and (3) the expression of IZUMO1 and JUNO during sperm maturation and folliculogenesis, respectively. Amplification for sequencing of feline IZUMO1 and JUNO was performed using the RT-PCR method. Levels of gene expression in different tissues were evaluated using real-time PCR. In situ hybridization was performed to localize JUNO mRNA in ovarian tissues. The complete coding sequences of IZUMO1 and JUNO were obtained and analyzed. A comparison between protein orthologs demonstrated the conservation of IZUMO1 and JUNO in Felidae. The real-time PCR results from various visceral organs indicated that IZUMO1 was significantly higher in the testis than in other organs, whereas JUNO was significantly higher in the ovary than in other organs. Expression of IZUMO1 was found to be higher in the testes than in the caput, corpus, and cauda of epididymides. In situ hybridization revealed that JUNO mRNA was in the ooplasm and nucleus of the primordial, primary, secondary, and antral follicles. Importantly, this was the first study to demonstrate the IZUMO1 and JUNO genes in the testis and ovary of cats. The results are useful for future research related to these genes and for developing contraceptives against these targets.
摘要：
由于手术绝育的耗时性质和家猫的快速繁殖速度，研究替代方案至关重要，该物种的非手术避孕方法。精子蛋白IZUMO1及其卵母细胞受体JUNO已被提议作为非手术避孕药的潜在靶标。本研究旨在证明（1）猫科动物IZUMO1和JUNO的蛋白质编码序列，(2)通过测定不同内脏组织mRNA水平,在特定器官中的基因表达,(3)IZUMO1和JUNO在精子成熟和卵泡发育过程中的表达,分别。使用RT-PCR方法进行用于猫科动物IZUMO1和JUNO的测序的扩增。使用实时PCR评估不同组织中的基因表达水平。进行原位杂交以定位卵巢组织中的JUNOmRNA。获得并分析了IZUMO1和JUNO的完整编码序列。蛋白质直向同源物之间的比较证明了IZUMO1和JUNO在Felidae中的保守性。各种内脏器官的实时PCR结果表明，睾丸中的IZUMO1明显高于其他器官，而JUNO在卵巢中明显高于其他器官。发现IZUMO1在睾丸中的表达高于在帽细胞中的表达，语料库，和附睾的尾囊。原位杂交显示JUNOmRNA存在于原始卵质和细胞核中，小学,次要,和窦卵泡。重要的是,这是第一项证明IZUMO1和JUNO基因在猫的睾丸和卵巢中的研究。该结果可用于与这些基因相关的未来研究以及开发针对这些靶标的避孕药。
