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The modulation of Smac/DIABLO on mitochondrial apoptosis induced by LPS in Crassostrea gigas
Lv, Zhao1,3; Song, Xiaorui2,4; Xu, Jiachao1,3; Jia, Zhihao1,3; Yang, Bin1,3; Jia, Yunke1,3; Qiu, Limei1; Wang, Lingling2,4,5; Song, Linsheng2,4,5
2019
Source PublicationFISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
Volume84Pages:587-598
Corresponding AuthorSong, Linsheng(lshsong@dlou.edu.cn)
AbstractThe mitochondrial pathway of apoptosis is well studied as the major mechanism of physiological cell death in vertebrates. In the present study, a second mitochondria-derived activator of caspases (Smac)/direct inhibitor of apoptosis-binding protein (IAP) with low pI protein (DIABLO) (designated as CgSmac) was identified from oyster Crassostrea gigas. The open reading frame of CgSmac was of 966 bp nucleotides encoding a predicted polypeptide of 321 amino acids with a conserved Smac/DIABLO domain containing a potential IAP-binding motif of VMPV. CgSmac proteins were distributed in hemocytes and co-localized with mitochondria. Western blotting analysis revealed that CgSmac proteins mainly existed in the dimer form in hemocytes, and the monomeric precursors and mature monomers were also detected. After lipopolysaccharide (LPS) stimulation, the mRNA expression of CgSmac in hemocytes was significantly up-regulated and peaked at 6 h (12.26-fold, p < 0.05), and the protein level of its dimers was significantly up-regulated at 6 h, 12 h, 24 h, and 48 h, while that of CgSmac monomers was up-regulated at 6 h, 12 h and down-regulated at 24 h, 48 h. The decrease of mitochondrial membrane potential indicated that the occurrence of early stage of apoptosis in primary cultured hemocytes was induced by LPS, and RNA interference (RNAi) of CgSmac could not rescue this decrease. The caspase-3 activity in primary cultured hemocytes was significantly suppressed after RNAi of CgSmac. Correspondingly, the total apoptotic rate of primary cultured hemocytes was also significantly suppressed in dsCgSmac + LPS group (31.57%) compared to dsEGFP + LPS group (40.27%, p < 0.05), which in turn demonstrated the conserved pro-apoptotic function of CgSmac. Furthermore, the early apoptotic rate (10.4% vs. 8.5%, p < 0.05) was significantly higher in dsCgSmac + LPS group than that of dsEGFP + LPS group, while the necrosis (7.7% vs. 10.0%, p < 0.05) and late apoptotic rates (13.4% vs. 21.9%, p < 0.05) were lower in dsCgSmac + LPS group than those of dsEGFP + LPS group. Collectively, CgSmac could activate mitochondrial apoptosis pathway by promoting caspase-3 activity in oyster hemocytes against exogenous LPS invasion. These results provided new insights on oyster apoptosis and the immune defense mechanisms in invertebrates.
KeywordMitochondrial pathway Smac/DIABLO LPS Primary cultured hemocytes C. gigas
DOI10.1016/j.fsi.2018.10.035
Indexed BySCI
Language英语
Funding ProjectNational Natural Science Foundation of China[31530069] ; Research Foundation for Talented Scholars in Dalian Ocean University ; AoShan Talents Cultivation Program - Qingdao National Laboratory for Marine Science and Technology[2017ASTCP-OS13] ; Research Foundation for Distinguished Professor in Liaoning ; Modern Agro-industry Technology Research System[CARS-49]
WOS Research AreaFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS SubjectFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS IDWOS:000457666800064
PublisherACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
Citation statistics
Cited Times:1[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/160865
Collection实验海洋生物学重点实验室
Corresponding AuthorSong, Linsheng
Affiliation1.Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China
2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266235, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
4.Dalian Ocean Univ, Liaoning Key Lab Marine Anim Immunol, Dalian 116023, Peoples R China
5.Dalian Ocean Univ, Liaoning Key Lab Marine Anim Immunol & Dis Contro, Dalian 116023, Peoples R China
First Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Recommended Citation
GB/T 7714
Lv, Zhao,Song, Xiaorui,Xu, Jiachao,et al. The modulation of Smac/DIABLO on mitochondrial apoptosis induced by LPS in Crassostrea gigas[J]. FISH & SHELLFISH IMMUNOLOGY,2019,84:587-598.
APA Lv, Zhao.,Song, Xiaorui.,Xu, Jiachao.,Jia, Zhihao.,Yang, Bin.,...&Song, Linsheng.(2019).The modulation of Smac/DIABLO on mitochondrial apoptosis induced by LPS in Crassostrea gigas.FISH & SHELLFISH IMMUNOLOGY,84,587-598.
MLA Lv, Zhao,et al."The modulation of Smac/DIABLO on mitochondrial apoptosis induced by LPS in Crassostrea gigas".FISH & SHELLFISH IMMUNOLOGY 84(2019):587-598.
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