Full Length Article
Expression profiling, immune functions, and molecular characteristics of the tetraspanin molecule CD63 from Amphiprion clarkii
-
- 발행기관
- 제주대학교 해양과학연구소 바로가기
-
- 간행물
- 해양과학연구소 연구논문집 바로가기
-
- 통권
- 제45권 (2021.12)바로가기
-
- 페이지
- pp.217-224
-
- 저자
-
- 언어
- 영어(ENG)
-
- URL
- https://www.earticle.net/Article/A406950
※ 기관로그인 시 무료 이용이 가능합니다.
원문정보
초록
- 영어
- CD63, a member of the tetraspanin family, is involved in the activation of immune cells, antiviral immunity, and signal transduction. The economically important anemonefishes Amphiprion sp. often face disease outbreaks, and the present study aimed to characterize CD63 in Amphiprion clarkii (denoted AcCD63) to enable better disease management. The in-silico analysis revealed that the AcCD63 transcript is 723 bp long and encodes 240 amino acids. The 26.2 kDa protein has a theoretical isoelectric point of 5.51. Similar to other tetraspanins, AcCD63 consists of four domains: short N-/C-terminal domains and small/large extracellular loops. Pairwise sequence alignment revealed that AcCD63 has the highest identity (100%) and similarity (99.2%) with CD63 from Amphiprion ocellaris. Multiple sequence alignment identified a conserved tetraspanin CCG motif, PXSCC motif, and C-terminal lysosome-targeting GYEVM motif. The quantitative polymerase chain reaction analysis showed that AcCD63 was highly expressed in the spleen and head kidney tissue, with low levels of expression in the liver. Temporal expression patterns of AcCD63 were measured in the head kidney and blood tissue after injection of polyinosinic:polycytidylic acid (poly (I:C)), lipolysacharides (LPS), or Vibrio harveyi (V. harveyi). AcCD63 was upregulated at 12 h post-injection with poly (I:C) or V. harveyi, and at 24 h post-injection with all stimulants in the head kidney. At 24 h post-injection, poly (I:C) and LPS upregulated, whereas V. harveyi downregulated AcCD63 expression in the blood. All viral hemorrhagic septicemia virus transcripts (M, G, N, RdRp, P, and NV) were downregulated in response to AcCD63 overexpression, and removal of viral particles occurred via the involvement of AcCD63. The expression of antiviral genes MX dynamin-like GTPase 1, interferon regulatory factor 3, interferon-stimulated gene 15, interferon-gamma, and viperin in CD63-overexpressing fathead minnow cells was downregulated. Collectively, our findings suggest that AcCD63 is an immunologically important gene involved in the A. clarkii pathogen stress response.
목차
ABSTRACT
1. Introduction
2. Materials and methods
2.1. AcCD63 sequence and bioinformatics analysis
2.2. Rearing of animals and challenge experiment
2.3. A. clarkii RNA extraction and cDNA synthesis
2.4. Spatial and temporal expression analysis
2.5. Plasmid construction for the viral assay
2.6. Cell culture conditions
2.7. Transfection of plasmids and viral infection
2.8. FHM cells and VHSV total RNA isolation and cDNA synthesis
2.9. qPCR of VHSV and FHM antiviral genes
2.10. Statistical analysis
3. Results
3.1. In-silico characterization of AcCD63
3.2. Spatial and temporal expression analysis
3.3. VHSV viral gene expression in AcCD63-overexpressing FHM cells
3.4. Mx, IRF3, ISG15, IFNγ, and Vip gene expression in AcCD63-overexpressing FHM cells
4. Discussion
References
키워드
CD63 Spatial and temporal expression VHSV Immune response Amphiprion clarkii저자
간행물 정보
발행기관
-
- 발행기관명
- 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]
- 간기
- 폐간
- pISSN
- 1225-5734
- 수록기간
- 1977~2021
- 십진분류
- KDC 454 DDC 551
이 권호 내 다른 논문 / 해양과학연구소 연구논문집 제45권
피인용수 : 0건 (자료제공 : 네이버학술정보)
함께 이용한 논문 이 논문을 다운로드한 분들이 이용한 다른 논문입니다.
0개의 논문이 장바구니에 담겼습니다.