Qiao Xue, Zhao Xinyu, Zeng Yuqing, Gu Xiaoyu, Wang Yiqing, Yu Hong, He Muchun, Wang Lingling, Song Linsheng
Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian, 116023, China; Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian, 116023, China; Dalian Key Laboratory of Aquatic Animal Disease Prevention and Control, Dalian Ocean University, Dalian, 116023, China.
Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian, 116023, China; Southern Laboratory of Ocean Science and Engineering, Zhuhai, Guangdong, Zhuhai, 519000, China; Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian, 116023, China; Dalian Key Laboratory of Aquatic Animal Disease Prevention and Control, Dalian Ocean University, Dalian, 116023, China.
Fish Shellfish Immunol. 2025 Aug;163:110363. doi: 10.1016/j.fsi.2025.110363. Epub 2025 Apr 21.
Rab5 functions as a pivotal regulator in the intricate processes of membrane trafficking, orchestrating a multitude of cellular activities. In the present study, a Rab5 homolog with conserved structure features was identified from Chinese mitten crab Eriocheir sinensis (designated EsRab5). The mRNA transcripts of EsRab5 were detected in all the tested tissues, with particularly high expression levels observed in brain and haematopoietic tissue (HPT). Notably, its mRNA expression in HPT was significantly up-regulated at 6 and 12 h following stimulation with Aeromonas hydrophila. Immunocytochemical assay showed that EsRab5 protein was diffusely distributed throughout the HPT, with a particularly prominent concentration in the cytoplasm. After A. hydrophila stimulation, the immunoreactive signals for EsRab5 in HPT were markedly more intense compared to those in the control group. Upon injection of EsRab5-specific siRNA to inhibit its expression, a significant increase in the percentage of EdU-positive cells within HPT was observed following A. hydrophila stimulation, which was 2.62-fold (p < 0.0001) of that in the EGFP-RNAi group. Meanwhile, the expression levels of proliferation related factors (EsRunx, EsGLP and EsAstakine), cell cycle-related proteins (EsCyclin E, EsCDK2, EsCDK4, and EsCyclin D) as well as the MAPK signal pathway were increased significantly in EsRab5-RNAi crabs after A. hydrophila stimulation. These results suggested that EsRab5 serves as a critical regulator in homeostasis maintenance of haematopoiesis in E. sinensis by modulating multiple factors.
Rab5在膜运输的复杂过程中作为关键调节因子发挥作用，协调众多细胞活动。在本研究中，从中华绒螯蟹（Eriocheir sinensis）中鉴定出一种具有保守结构特征的Rab5同源物（命名为EsRab5）。在所有测试组织中均检测到EsRab5的mRNA转录本，在脑和造血组织（HPT）中观察到特别高的表达水平。值得注意的是，在嗜水气单胞菌刺激后6小时和12小时，其在HPT中的mRNA表达显著上调。免疫细胞化学分析表明，EsRab5蛋白在整个HPT中呈弥漫性分布，在细胞质中浓度尤为突出。嗜水气单胞菌刺激后，HPT中EsRab5的免疫反应信号明显强于对照组。注射EsRab5特异性siRNA抑制其表达后，嗜水气单胞菌刺激后HPT中EdU阳性细胞的百分比显著增加，是EGFP-RNAi组的2.62倍（p < 0.0001）。同时，嗜水气单胞菌刺激后，EsRab5-RNAi蟹中增殖相关因子（EsRunx、EsGLP和EsAstakine）、细胞周期相关蛋白（EsCyclin E、EsCDK2、EsCDK4和EsCyclin D）以及MAPK信号通路的表达水平显著增加。这些结果表明，EsRab5通过调节多种因素在中华绒螯蟹造血稳态维持中起关键调节作用。
