Molecular mechanism of LncRNA regulating meiotic chromosome axis assembly through phase separation in rabbit spermatogenesis
LncRNA通过相分离调控减数分裂染色体轴组装在家兔精子发生中的分子机制

摘要

目的 研究长链非编码RNA（LncRNA）通过相分离调控减数分裂染色体轴组装在家兔精子发生中的分子机制。方法 采用健康雄性新西兰白兔开展研究，进行睾丸样本采集，进行LncRNA筛选、鉴定、相分离能力验证、染色体轴组装检测等。结果 家兔减数分裂各阶段共鉴定出2847个LncRNA，符合差异表达标准的LncRNA中，减数分裂不同阶段的阶段上调、LncRNA数量、特异性高表达LncRNA、染色体轴基因共表达LncRNA等存在差异。LncRNA-XY1敲减实验中，家兔精母细胞的SYCP3轴长显著缩短，完整SC比例、同源染色体配对率显著降低，减数分裂停滞率显著升高。相较于普通家兔，LncRNA敲减家兔精子畸形率显著升高，前向运动精子比例、精子平均路径速度、顶体反应率显著降低。结论 LncRNA表达缺失会导致染色体轴组装异常、减数分裂阻滞，诱发精子畸形率的升高与活力的下降。

Abstract

Objective To study the molecular mechanism of long non coding RNA (LncRNA) regulating meiotic chromosome axis assembly in rabbit spermatogenesis through phase separation.
Methods healthy male New Zealand white rabbits were used to carry out the study. Testicular samples were collected for LncRNA screening, identification, phase separation ability verification, chromosome axis assembly detection, etc.
Results a total of 2847 LncRNA were identified at each stage of meiosis in rabbits. Among the LncRNA that met the differential expression criteria, there were differences in the stage of meiosis, the number of LncRNA, the specific high expression of LncRNA, and the co expression of chromosome axis genes. In LncRNA-XY1 knockdown test, the SYCP3 axis length of rabbit spermatocytes was significantly shortened, the proportion of complete SC and homologous chromosome pairing rate were significantly reduced, and the meiotic arrest rate was significantly increased. Compared with normal rabbits, the sperm deformity rate of LncRNA knockout rabbits was significantly increased, and the proportion of forward motile sperm, average sperm path velocity and acrosome reaction rate were significantly decreased.
Conclusion   the loss of LncRNA expression can lead to abnormal assembly of chromosome axis, meiosis block, and the increase of sperm deformity rate and the decrease of sperm motility.