Molecular insights and immune responses of big belly seahorse syndecan-2 (CD362): Involvement of ectodomain in regulating cell survival, proliferation, and wound healing
Syndecan-2, also known as CD362, is a transmembrane heparan sulfate proteoglycan which regulates cell growth, proliferation, cell adhesion, wound healing, and recruits immune cells. In the present study, we performed bioinformatics, spatial and temporal expression analyses of Hippocampus abdominalis syndecan-2 (HaSDC-2). Additionally, functional assays were conducted. HaSDC-2 has five major domains; an extracellular heparan sulfate attachment domain, a co-receptor binding domain, a transmembrane domain, two conserved domains (C1 domain, C2 domain), and a variable (V) domain. The ectodomain contained a signal peptide and GAG attachment sites. In-silico analysis revealed that HaSDC-2 contained a 798 bp long ORF and protein sequence of 265 amino acid residues. Further analysis of the amino acid sequence predicted a 28.9 kDa molecular weight and a 4.13 theoretical isoelectric point. The spatial expression of HaSDC-2 was ubiquitous in all tested tissues. HaSDC-2 expression in the liver was upregulated 24 h post-injection in response to all stimuli. Further, HaSDC-2 expression in blood cells was upregulated at 12 and 72 h post-injection in response to all the stimuli. HaSDC-2 + pcDNA™3.1(+) transfected cells exhibited significant survival in response to cell stressors such as H2O2 and HED. The ectodomain of recombinant HaSDC-2 treated cells showed significant cell proliferation in a concentration-dependent manner. The scratch wound healing assay showed significant Δ gap closures with increasing concentrations of HaSDC-2. Collectively, these results indicated that syndecan-2 was involved in regulating immune responses and cell stress conditions.
목차
ABSTRACT 1. Introduction 2. Materials and methods 2.1. Identification of HaSDC-2 sequence and characterization 2.2. Rearing of fish, immune challenge and isolation of tissues 2.3. Total RNA extraction and cDNA synthesis 2.4. Spatial and temporal expression analysis 2.5. Preparation of plasmids, cell culture maintenance, and transfection 2.6. Cellular toxicity/viability assay and cell proliferation assay 2.7. Scratch wound healing assay 3. Results and discussion 3.1. Sequence characterization 3.2. Spatial and temporal expression analysis 3.3. HaSDC-2 overexpression in FHM cells leads to cell survival 3.4. Effect of recombinant HaSDC-2-ED on cell proliferation 3.5. Recombinant HaSDC-2-ED in wound healing 4. Conclusion Acknowledgement References
키워드
Cell viability and proliferationImmune challengeSDC-2Seahorse
저자
D.S. Liyanage [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea ]
W.K.M. Omeka [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea ]
M.D. Neranjan Tharuka [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
Sumi Jung [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
Sukkyoung Lee [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
Jehee Lee [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
Corresponding Author
제주대학교 해양과학연구소 [Marine Science Institute Jeju National University]
설립연도
1968
분야
농수해양>수산학
소개
해양의 실체와 그 자원의 합리적 이용과 관리방법을 규명하기 위하여 해양물리, 화학, 생물, 지질, 공학등 여러 관련분야에 관한 다양한 연구를 수행하며, 환경의 보전과 보호를 위해서 수질, 대기, 상·하수도, 폐기물, 사회적 환경 등의 연구를 통하여 쾌적하고 청정한 환경을 보전하기 위한 다양한 연구를 수행하고, 또한 해양 및 환경분야의 우수한 인력양성, 국내외 학술교류의 증진, 학·산 협동체제의 확립 등을 통하여 해양과 환경에 관한 기초 및 응용적 학술연구를 종합적으로 수행함을 목적으로 한다.
간행물
간행물명
해양과학연구소 연구논문집 [Bulletin of the Marine Science Institute]