Yan Zhi-Jun, Wu Wen-Tao, Xu Lian-Ying, Bi Ning, Yang Fan, Yang Wei-Jun, Yang Jin-Shu
College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 3100058, China.
College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 3100058, China.
Gene. 2023 May 25;866:147349. doi: 10.1016/j.gene.2023.147349. Epub 2023 Mar 8.
To survive under harsh environments, embryonic development of Artemia was arrested at the gastrula stage and released as the diapause embryo. Cell cycle and metabolism were highly suppressed in this state of quiescence. However, cellular mechanisms underlying diapause remain largely unclear. In this study, we found that the expression level of a CT10 regulator of kinase-encoding gene (Ar-Crk) in diapause embryos was significantly lower than non-diapause embryos at the early embryogenetic stage of Artemia. Knockdown of Ar-Crk by RNA interference induced formation of diapause embryos, while the control group produced nauplii. Western blot analysis and metabolic assays revealed that the diapause embryos produced by Ar-Crk-knocked-down Artemia had similar characteristics of diapause markers, arrested cell cycle, and suppressed metabolism with those diapause embryos produced by natural oviparous Artemia. Transcriptomic analysis of Artemia embryos revealed knockdown of Ar-Crk induced downregulation of the aurora kinase A (AURKA) signaling pathway, as well as energetic and biomolecular metabolisms. Taken together, we proposed that Ar-Crk is a crucial factor in determining the process of diapause in Artemia. Our results provide insight into the functions of Crk in fundamental regulations such as cellular quiescence.
为了在恶劣环境中生存，卤虫的胚胎发育在原肠胚阶段停滞，并以滞育胚胎的形式释放。在这种静止状态下，细胞周期和新陈代谢受到高度抑制。然而，滞育背后的细胞机制在很大程度上仍不清楚。在本研究中，我们发现卤虫滞育胚胎中一种编码激酶的CT10调节基因（Ar-Crk）的表达水平在胚胎发育早期显著低于非滞育胚胎。通过RNA干扰敲低Ar-Crk诱导了滞育胚胎的形成，而对照组产生了无节幼体。蛋白质免疫印迹分析和代谢测定表明，经Ar-Crk敲低的卤虫产生的滞育胚胎与自然产卵卤虫产生的滞育胚胎具有相似的滞育标记特征、细胞周期停滞和代谢抑制。卤虫胚胎的转录组分析表明，敲低Ar-Crk会导致极光激酶A（AURKA）信号通路以及能量和生物分子代谢的下调。综上所述，我们提出Ar-Crk是决定卤虫滞育过程的关键因素。我们的结果为Crk在细胞静止等基本调节中的功能提供了见解。
