在太平洋牡蛎中，CgATP合酶β亚基作为一种CgAstakine受体参与血细胞增殖的调控。
CgATP synthase β subunit involved in the regulation of haemocytes proliferation as a CgAstakine receptor in Crassostrea gigas.
作者信息
Yang Ying, Qiao Xue, Song Xiaorui, Zhang Dan, Yu Simiao, Dong Miren, Liu Xiyang, Wang Lingling, Song Linsheng
机构信息
Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian, 116023, China; 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; Functional Laboratory of Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266235, China; Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian, 116023, China; Dalian Key Laboratory of Aquatic Animal Disease Prevention and Control, Dalian Ocean University, Dalian, 116023, China.
出版信息
Fish Shellfish Immunol. 2022 Apr;123:85-93. doi: 10.1016/j.fsi.2022.02.054. Epub 2022 Mar 2.
Astakine is considered as an endogenous cytokine-like factor of prokineticin homologue in invertebrate. Recently, an astakine homologue (CgAstakine) has been identified and characterized in oyster Crassostrea gigas. In the present study, a CgATP synthase β subunit was identified as the receptor of CgAstakine in C. gigas. There was an ATP-synt_ab_N domain and an AAA domain in the CgATP synthase β subunit protein. The mRNA transcripts of CgATP synthase β subunit were detected in all tested tissues, with the highest expression level in hepatopancreas and gills, which was 109.11-fold (p < 0.01) and 97.21-fold (p < 0.01) of that in labial palps, respectively. After rCgAstakine stimulation, the mRNA transcripts of CgATP synthase β subunit in agranulocytes and semi-granulocytes were significantly increased at 24 h (2.44-fold, and 9.01-fold of that in control group, p < 0.01), and those in granulocytes were significantly increased at 6 h (1.83-fold, p < 0.01), 12 h (1.92-fold, p < 0.01) and 24 h (3.47-fold, p < 0.01). The expression level of CgATP synthase β subunit protein in agranulocytes and granulocytes was also significantly increased after rCgAstakine stimulation, which was 1.64-fold (p < 0.05) and 1.85-fold (p < 0.05) of that in control group, respectively, while there were no significant changes in semi-granulocytes. The immunofluorescence assay showed that CgATP synthase β subunit positive signals were mainly located on the membrane of haemocytes. The number of haemocytes with EdU positive signals was significantly increased after rCgAstakine stimulation (2.04-fold of seawater group, p < 0.01), while significantly decreased after the RNA interference (RNAi) of CgATP synthase β subunit, which was 0.28-fold of that in NC group (p < 0.01). Bio-layer interferometry (BLI) assay confirmed in vitro interaction between rCgAstakine and rCgATP synthase β subunit. There results suggested that CgATP synthase β subunit acts as the receptor of CgAstakine and plays important roles in CgAstakine induced renewal of haemocytes in C. gigas.
促动素被认为是无脊椎动物中前动力蛋白同源物的一种内源性细胞因子样因子。最近，在太平洋牡蛎中鉴定并表征了一种促动素同源物（CgAstakine）。在本研究中，鉴定出太平洋牡蛎中的CgATP合酶β亚基为CgAstakine的受体。CgATP合酶β亚基蛋白中有一个ATP - synt_ab_N结构域和一个AAA结构域。在所有测试组织中均检测到CgATP合酶β亚基的mRNA转录本，在肝胰腺和鳃中的表达水平最高，分别是唇瓣中表达水平的109.11倍（p < 0.01）和97.21倍（p < 0.01）。rCgAstakine刺激后，无颗粒细胞和半颗粒细胞中CgATP合酶β亚基的mRNA转录本在24小时时显著增加（分别是对照组的2.44倍和9.01倍，p < 0.01），而颗粒细胞中的mRNA转录本在6小时（1.83倍，p < 0.01）、12小时（1.92倍，p < 0.01）和24小时（3.47倍，p < 0.01）时显著增加。rCgAstakine刺激后，无颗粒细胞和颗粒细胞中CgATP合酶β亚基蛋白的表达水平也显著增加，分别是对照组的1.64倍（p < 0.05）和1.85倍（p < 0.05），而半颗粒细胞中无显著变化。免疫荧光分析表明，CgATP合酶β亚基阳性信号主要位于血细胞的膜上。rCgAstakine刺激后，EdU阳性信号的血细胞数量显著增加（是海水组的2.04倍，p < 0.01），而CgATP合酶β亚基的RNA干扰（RNAi）后显著减少，是NC组的0.28倍（p < 0.01）。生物层干涉术（BLI）分析证实了rCgAstakine与rCgATP合酶β亚基之间的体外相互作用。这些结果表明，CgATP合酶β亚基作为CgAstakine的受体，在CgAstakine诱导的太平洋牡蛎血细胞更新中起重要作用。
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