CgCaspase-3 激活太平洋牡蛎血细胞中 CgGSDME 的易位。
CgCaspase-3 activates the translocation of CgGSDME in haemocytes of Pacific oyster Crassostrea gigas.
机构信息
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.
Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian, 116023, China; Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Zhuhai, 519000, 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 Dec;131:757-765. doi: 10.1016/j.fsi.2022.10.036. Epub 2022 Oct 21.
Cysteinyl aspartate specific proteinase-3 (Caspase-3) is an important protein involved in the apoptosis and gasdermin E (GSDME)-mediated cell pyroptosis pathways in vertebrates. A Caspase-3 homologue (designated as CgCaspase-3) was previously identified as an immune receptor specific for lipopolysaccharide (LPS) to regulate apoptosis in the Pacific oyster Crassostrea gigas. In the present study, the binding activity of CgCaspase-3 to different pathogen associated molecular patterns (PAMPs) and its effects on CgGSDME translocation in haemocytes were further investigated in C. gigas. The mRNA expression of CgCaspase-3 could be detected in all the tested tissues, including hepatopancreas, labial palp, adductor muscle, gonad, gill, mantle and haemocytes, and it was highly expressed in labial palp, gonad, haemocytes, and adductor muscle. The mRNA expression of CgCaspase-3 in haemocytes increased significantly at 3, 24, 48 and 72 h after LPS stimulation, and it increased significantly at 6, 12, 24 and 48 h after Vibrio splendidus stimulation. The recombinant CgCaspase-3 displayed binding activity towards LPS, mannose (MAN), peptidoglycan (PGN), and polyinosinic-polycytidylic acid potassium salt (Poly (I:C)). The positive signals of CgGSDME on haemocyte membrane became stronger at 3 h after V. splendidus stimulation, compared with that of Seawater group, and the co-localization of CgCaspase-3 and CgGSDME was observed in the haemocyte membrane. After the injection of dsCgCaspase-3, the positive signals of CgGSDME on haemocyte membrane became weaker compared with that of EGFP-RNAi group at 24 h after V. splendidus stimulation. The results suggested that CgCaspase-3 was able to bind diverse PAMPs and activate the translocation of CgGSDME in haemocytes of oyster response against pathogen invasion.
半胱氨酸天冬氨酸特异性蛋白酶-3（Caspase-3）是脊椎动物细胞凋亡和gasdermin E（GSDME）介导的细胞焦亡途径中的重要蛋白。先前已经鉴定出 Caspase-3 同源物（命名为 CgCaspase-3）是一种对脂多糖（LPS）具有特异性的免疫受体，用于调节太平洋牡蛎 Crassostrea gigas 的细胞凋亡。在本研究中，进一步研究了 C. gigas 中 CgCaspase-3 与不同病原体相关分子模式（PAMPs）的结合活性及其对 CgGSDME 向血细胞中转位的影响。在所有测试的组织中，包括肝胰腺、唇瓣、闭壳肌、性腺、鳃、套膜和血细胞，都可以检测到 CgCaspase-3 的 mRNA 表达，并且在唇瓣、性腺、血细胞和闭壳肌中表达水平较高。在 LPS 刺激后 3、24、48 和 72 h，血细胞中 CgCaspase-3 的 mRNA 表达显著增加，在 Vibrio splendidus 刺激后 6、12、24 和 48 h 也显著增加。重组 CgCaspase-3 对 LPS、甘露糖（MAN）、肽聚糖（PGN）和聚肌苷酸-聚胞苷酸钾盐（Poly（I：C））表现出结合活性。与海水组相比，在 V. splendidus 刺激后 3 h，血细胞膜上的 CgGSDME 阳性信号变强，并且在血细胞膜上观察到 CgCaspase-3 和 CgGSDME 的共定位。在注射 dsCgCaspase-3 后，与 EGFP-RNAi 组相比，在 V. splendidus 刺激后 24 h，血细胞膜上的 CgGSDME 阳性信号变弱。结果表明，CgCaspase-3 能够结合多种 PAMPs，激活牡蛎血细胞对病原体入侵的反应中 CgGSDME 的易位。
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