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The activated beta-integrin (Cg beta V) enhances RGD-binding and phagocytic capabilities of hemocytes in Crassostrea gigas
Lv, Zhao1,2,3; Qiu, Limei1; Jia, Zhihao1,3; Wang, Weilin2,4; Liu, Zhaoqun4,5; Wang, Lingling4,5; Song, Linsheng2,4,5
2019-04-01
Source PublicationFISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
Volume87Pages:638-649
Corresponding AuthorSong, Linsheng(lshsong@dlou.edu.cn)
AbstractIntegrins are an important family of cell receptors that can bind foreign particles and promote cell phagocytosis after they are activated. In the present study, a novel beta integrin was identified from pacific oyster Crassostrea gigas with an extracellular domain, a single transmembrane segment, and a short cytoplasmic domain. It was phylogenetically clustered with phagocytosis-related insecta beta V, and designated as Cg beta V. Cg beta V shared a conserved NPX[Y/F] motif related to integrin activation with other phagocytosis-related beta integrins. The mRNA transcripts of Cg beta V were widely detected in oyster tissues including hemocytes, gonad, adductor muscle, mantle, gill, and hepatopancreas, and the expression level in hemocytes was significantly up-regulated at 12 h after lipopolysaccharide (LPS) stimulation (p < 0.05), which was 2.29-fold higher than that in the control group. Cg beta V proteins were mainly observed on the hemocytes surface. The oyster hemocytes were found to bind fluorescein isothiocyanate (FITC)-labeled Arg-Gly-Asp-containing peptides (RGDCPs), and the binding capability was significantly up-regulated with the peak percentage of 37.6% at 12 h post LPS stimulation, which was higher than that in the control group (8.8%, p < 0.05), suggesting the activation of RGD-binding integrins on oyster hemocytes surface. The label-free RGDCPs and anti-Cg beta V antibody inhibited the binding capability of hemocytes towards FITC-labeled RGDCPs, which were significant lower in RGD blocking group (7.4%, p < 0.05) and anti-Cg beta V blocking group (22.1%, p < 0.05) than that in the control group (37.6%), indicating that Cg beta V could be a RGD-binding integrin. Phagocytosis assay demonstrated that LPS could enhance the phagocytosis of hemocytes towards Escherichia cob and fluorescent beads with the phagocytic rate (PR) of 18.3% and 17.4%, and phagocytic index (PI) of 5.29 and 37.71, respectively, which were significant higher than that in the control group (11.0% and 3.65 for E. coli, 9.8% and 29.26 for fluorescent beads, respectively, p < 0.05). In addition, both the label-free RGDCPs and anti-Cg beta V antibody significantly hindered the phagocytosis of hemocytes towards E. coli and fluorescent beads. After the E. coli and fluorescent beads were opsonized by oyster serum, the label-free RGDCPs still inhibited the phagocytosis of hemocytes towards them, while the anti-Cg beta V antibody could only inhibit the phagocytosis of hemocytes towards E. coli, suggesting that only the activated Cg beta V was involved in the enhancing phagocytosis for bacteria in oyster. Moreover, the key components of conserved integrin-mediated phagocytosis pathway including GTPases, talin proteins, Ca2+ and cAMP were all induced by LPS in hemocytes of oyster. All these results suggested that the activated Cg beta V enhanced RGD-binding and phagocytic capabilities of hemocytes, shedding lights on the mechanisms of integrin-mediated phagocytosis in mollusks.
KeywordC. gigas RGD-Binding Integrin activation Integrin-mediated phagocytosis
DOI10.1016/j.fsi.2019.01.047
Indexed BySCI
Language英语
Funding ProjectNational Natural Science Foundation of China[U1706204] ; National Natural Science Foundation of China[31530069] ; Modern Agroindustry Technology Research System[CARS-49] ; AoShan Talents Cultivation Program by Qingdao National Laboratory for Marine Science and Technology[2017ASTCP-OS13] ; Dalian High Level Talent Innovation Support Program[2015R020] ; Research Foundation for Talented Scholars in Dalian Ocean University
WOS Research AreaFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS SubjectFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS IDWOS:000464086600070
PublisherACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
Citation statistics
Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/161138
Collection实验海洋生物学重点实验室
Corresponding AuthorSong, Linsheng
Affiliation1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Shandong, Peoples R China
2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266235, Shandong, 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,Qiu, Limei,Jia, Zhihao,et al. The activated beta-integrin (Cg beta V) enhances RGD-binding and phagocytic capabilities of hemocytes in Crassostrea gigas[J]. FISH & SHELLFISH IMMUNOLOGY,2019,87:638-649.
APA Lv, Zhao.,Qiu, Limei.,Jia, Zhihao.,Wang, Weilin.,Liu, Zhaoqun.,...&Song, Linsheng.(2019).The activated beta-integrin (Cg beta V) enhances RGD-binding and phagocytic capabilities of hemocytes in Crassostrea gigas.FISH & SHELLFISH IMMUNOLOGY,87,638-649.
MLA Lv, Zhao,et al."The activated beta-integrin (Cg beta V) enhances RGD-binding and phagocytic capabilities of hemocytes in Crassostrea gigas".FISH & SHELLFISH IMMUNOLOGY 87(2019):638-649.
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