PMID- 33275994 OWN - NLM STAT- MEDLINE DCOM- 20220126 LR - 20220126 IS - 1879-0089 (Electronic) IS - 0145-305X (Linking) VI - 116 DP - 2021 Mar TI - Hemocyte extracellular traps of Manila clam Ruditapes philippinarum: Production characteristics and antibacterial effects. PG - 103953 LID - S0145-305X(20)30508-5 [pii] LID - 10.1016/j.dci.2020.103953 [doi] AB - Extracellular traps (ETs) have been found to be an important strategy of mammals to immobilize and kill invading microorganisms. In the present study, we observed the formation of ETs in the hemocytes of marine mollusks Ruditapes philippinarum in response to challenge from bacteria Vibrio anguillarum, and examined the potential factors and signaling pathways underling this process. We detected an increase of reactive oxygen species (ROS) and myeloperoxidase (MPO) production during ETosis, accompanied by significantly up-regulated expression of ROS-related and MPO genes. The suppression of ETs structures by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor (diphenyleneiodonium chloride, DPI) and MPO inhibitor (aminobenzoic acid hydrazide, ABAH) further confirmed the essential roles ROS and MPO played in ETosis. Furthermore, ET production could be inhibited by phosphotidylinsitol-3-kinase (PI3K) inhibitor (LY294002) and extracellular regulated protein kinase (ERK) inhibitor (U0126), suggesting the idea that both the PI3K and ERK pathways were suggested to function during ETosis. In addition, the ETosis process was accompanied by enhancement of glycolysis-related enzymatic activities, e.g., pyruvate kinase (PK) and hexokinase (HK), and over-expression of the glycolysis-related genes, e.g., PK, HK and glucose transport protein (GLUT), indicating high involvement of glycolysis in the ETosis process. Furthermore, our scanning electron microscopy (SEM) observation and antibacterial activities test successfully showed the patterns how clam ETs entrapped and killed the invading V. anguillarum. Taken together, our results revealed that ETosis with bactericidal effect increased ROS, MPO and glycolysis level and carried out in a ROS-, MPO-, PI3K-ERK-dependent manner. CI - Copyright (c) 2020 Elsevier Ltd. All rights reserved. FAU - Han, Yijing AU - Han Y AD - Muping Coastal Environment Research Station, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, Shandong, 264003, PR China; Research and Development Center for Efficient Utilization of Coastal Bioresources, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, Shandong, 264003, PR China; Center for Ocean Mega-science, Chinese Academy of Sciences, Qingdao, Shandong, 266071, PR China; University of Chinese Academy of Sciences, Beijing, 100049, PR China. FAU - Chen, Lizhu AU - Chen L AD - Shandong Marine Resource and Environment Research Institute, Yantai, 264006, PR China. FAU - Zhang, Qianqian AU - Zhang Q AD - Muping Coastal Environment Research Station, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, Shandong, 264003, PR China; Research and Development Center for Efficient Utilization of Coastal Bioresources, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, Shandong, 264003, PR China; Center for Ocean Mega-science, Chinese Academy of Sciences, Qingdao, Shandong, 266071, PR China. FAU - Yu, Daode AU - Yu D AD - Marine Biology Institute of Shandong Province, Qingdao, Shandong, 266002, PR China. FAU - Yang, Dinglong AU - Yang D AD - Muping Coastal Environment Research Station, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, Shandong, 264003, PR China; Research and Development Center for Efficient Utilization of Coastal Bioresources, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, Shandong, 264003, PR China; Center for Ocean Mega-science, Chinese Academy of Sciences, Qingdao, Shandong, 266071, PR China. Electronic address: dlyang@yic.ac.cn. FAU - Zhao, Jianmin AU - Zhao J AD - Muping Coastal Environment Research Station, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, Shandong, 264003, PR China; Research and Development Center for Efficient Utilization of Coastal Bioresources, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, Shandong, 264003, PR China; Center for Ocean Mega-science, Chinese Academy of Sciences, Qingdao, Shandong, 266071, PR China. Electronic address: jmzhao@yic.ac.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20201201 PL - United States TA - Dev Comp Immunol JT - Developmental and comparative immunology JID - 7708205 RN - 0 (Enzyme Inhibitors) RN - 0 (Reactive Oxygen Species) RN - EC 1.11.1.7 (Peroxidase) RN - Vibrio anguillarum SB - IM MH - Animals MH - Bivalvia/*immunology/microbiology MH - Enzyme Inhibitors/pharmacology MH - Extracellular Traps/drug effects/*immunology/metabolism/microbiology MH - Glycolysis MH - Hemocytes/*immunology/metabolism/microbiology MH - Immunity, Innate/genetics MH - Microbial Viability MH - Peroxidase/antagonists & inhibitors/metabolism MH - Reactive Oxygen Species/antagonists & inhibitors/metabolism MH - Signal Transduction/drug effects MH - Vibrio/physiology OTO - NOTNLM OT - Bactericidal effect OT - Extracellular traps OT - Glycolysis OT - Reactive oxygen species OT - Ruditapes philippinarum EDAT- 2020/12/05 06:00 MHDA- 2022/01/27 06:00 CRDT- 2020/12/04 20:08 PHST- 2020/08/31 00:00 [received] PHST- 2020/11/27 00:00 [revised] PHST- 2020/11/28 00:00 [accepted] PHST- 2020/12/05 06:00 [pubmed] PHST- 2022/01/27 06:00 [medline] PHST- 2020/12/04 20:08 [entrez] AID - S0145-305X(20)30508-5 [pii] AID - 10.1016/j.dci.2020.103953 [doi] PST - ppublish SO - Dev Comp Immunol. 2021 Mar;116:103953. doi: 10.1016/j.dci.2020.103953. Epub 2020 Dec 1.