Formation of different polyploids through disrupting meiotic crossover frequencies based on cntd1 knockout in zebrafish

• Published in: Molecular biology and evolution 2024-03, Vol.41 (3)
• Main Authors: Ou, Yuan, Li, Huilin, Li, Juan, Dai, Xiangyan, He, Jiaxin, Wang, Shi, Liu, Qingfeng, Yang, Conghui, Wang, Jing, Zhao, Rurong, Yin, Zhan, Shu, Yuqin, Liu, Shaojun
• Format: Article
• Language: English
• Subjects: Discoveries
• ISSN: 0737-4038; 1537-1719
• DOI: 10.1093/molbev/msae047

Polyploidy, a significant catalyst for speciation and evolutionary processes in both plant and animal kingdoms, has been recognized for a long time. However, the exact molecular mechanism that leads to polyploid formation, especially in vertebrates, is not fully understood. Our study aimed to elucidate this phenomenon using the zebrafish model. We successfully achieved an effective knockout of the cyclin N-terminal domain containing 1 (cntd1) using CRISPR/Cas9 technology. This resulted in impaired formation of meiotic crossovers, leading to cell-cycle arrest during meiotic metaphase and triggering apoptosis of spermatocytes in the testes. Despite these defects, the mutant (cntd1-/-) males were still able to produce a limited amount of sperm with normal ploidy and function. Interestingly, in the mutant females, it was the ploidy, not the capacity of egg production that was altered. This resulted in the production of haploid, aneuploid, and unreduced gametes. This alteration enabled us to successfully obtain triploid and tetraploid zebrafish from cntd1-/- and cntd1-/-/- females, respectively. Furthermore, the tetraploid-heterozygous zebrafish produced reduced-diploid gametes and yielded all-triploid or all-tetraploid offspring when crossed with wild-type (WT) or tetraploid zebrafish, respectively. Collectively, our findings provide direct evidence supporting the crucial role of meiotic crossover defects in the process of polyploidization. This is particularly evident in the generation of unreduced eggs in fish and, potentially, other vertebrate species.