Cite this paper:
LI Zan, LIU Xiumei, DU Xinxin, ZHANG Kai, CHEN Yan, WANG Xubo, WANG Zhigang, YU Haiyang, ZHANG Quanqi. Tumor necrosis factor-alpha (TNF-α) in spotted halibut Verasper variegatus at the embryonic and metamorphic stages[J]. HaiyangYuHuZhao, 2020, 38(2): 454-466

Tumor necrosis factor-alpha (TNF-α) in spotted halibut Verasper variegatus at the embryonic and metamorphic stages

LI Zan1,3, LIU Xiumei2, DU Xinxin3, ZHANG Kai3, CHEN Yan3, WANG Xubo3, WANG Zhigang3, YU Haiyang3, ZHANG Quanqi3
1 School of Agriculture, Ludong University, Yantai 264025, China;
2 College of Life Sciences, Yantai University, Yantai 264005, China;
3 Key Laboratory of Marine Genetics and Breeding, Ministry of Education, College of Marine Life Science, Ocean University of China, Qingdao 266003, China
As an aquatic fish, the spotted halibut Verasper variegatus is highly susceptible to bacterial and virus infections. Tumor necrosis factor-alpha (TNF-α) as a cytokine could control the inflammatory responses. The functions of TNF-α in many species have been widely studied, particularly in mammals. However, little is known about the TNF-α functions in V. variegatus. We first cloned and sequenced the TNF-α gene in V. variegatus (VvTNF-α). The two conserved cysteine residues, transmembrane sequence, Thr-Leu motif, and TNF family signature, as well as the TA-rich motifs of its proteins related to inflammatory responses had high similarity to those of the other teleost and mammalian TNF-α. The phylogenetic analysis showed that VvTNF-α was consistent with TNF-α genes of other vertebrates. The VvTNF-α transcripts were extensively distributed in the peripheral blood leukocytes (PBLs), spleen, and gill, indicating that the VvTNF-α had a role in immune function. Furthermore, treatment with pathogen-associated molecular patterns (PAMPs) could induce a rapid and significant increase of VvTNF-α in the PBLs, which reveals that VvTNF-α does participate in the host immune responses against bacterial and viral pathogens. We found that VvTNF-α had an interesting expression pattern during metamorphosis, showing that the flatfish TNF-α may have some novel functions during specific developmental stages. In addition, the 3D structure prediction of VvTNF-α provided an indication of how it is likely to interact with other proteins. Therefore, VvTNF-α has multiple functions, and provides valuable information to explore novel functions of TNF-α.
Key words:    tumor necrosis factor-alpha (TNF-α)|Verasper variegatus|metamorphosis|peripheral blood leukocytes (PBLs)|3D modeling|pathogen-associated molecular patterns (PAMPs)   
Received: 2018-11-08   Revised: 2019-01-28
PDF (1146 KB) Free
Print this page
Add to favorites
Email this article to others
Articles by LI Zan
Articles by LIU Xiumei
Articles by DU Xinxin
Articles by ZHANG Kai
Articles by CHEN Yan
Articles by WANG Xubo
Articles by WANG Zhigang
Articles by YU Haiyang
Articles by ZHANG Quanqi
Acton R D, Dahlberg P S, Uknis M E, Klaerner H G, Fink G S, Norman J G, Dunn D L. 1996. Differential sensitivity to Escherichia coli infection in mice lacking tumor necrosis factor p55 or interleukin-1 p80 receptors. Archives of Surgery, 131(11):1 216-1 221.
Aust G, Heuer M, Laue S, Lehmann I, Hofmann A, Heldin N E, Scherbaum W A. 1996. Expression of tumour necrosis factor-alpha (TNF-α) mRNA and protein in pathological thyroid tissue and carcinoma cell lines. Clinical and Experimental Immunology, 105(1):148-154.
Beg A A, Baltimore D. 1996. An essential role for NF-κB in preventing TNF-α-induced cell death. Science, 274(5288):782-784.
Blobel C P. 1997. Metalloprotease-disintegrins:links to cell adhesion and cleavage of TNFα and Notch. Cell, 90(4):589-592.
Campinho M A, Silva N, Sweeney G E, Power D M. 2007. Molecular, cellular and histological changes in skin from a larval to an adult phenotype during bony fish metamorphosis. Cell and Tissue Research, 327(2):267-284.
Camussi G, Albano E, Tetta C, Bussolino F. 1991. The molecular action of tumor necrosis factor-α. European Journal of Biochemistry, 202(1):3-14.
Caput D, Beutler B, Hartog K, Thayer R, Brown-Shimer S, Cerami A. 1986. Identification of a common nucleotide sequence in the 3'-untranslated region of mRNA molecules specifying inflammatory mediators. Proceedings of the National Academy of Sciences of the United States of America, 83(6):1 670-1 674.
Chang M X, Nie P, Xie H X, Wang G L, Gao Y. 2006.Characterization and expression analysis of TNF-related apoptosis inducing ligand (TRAIL) in grass carp Ctenopharyngodon idella. Veterinary Immunology and Immunopathology, 110(1-2):51-63.
Chu W. 2013. Tumor necrosis factor. Cancer Letters, 328(2):222-225.Covello J M, Bird S, Morrison R N, Battaglene S C, Secombes C J, Nowak B F. 2009. Cloning and expression analysis of three striped trumpeter (Latris lineata) pro-inflammatory cytokines, TNF-α, IL-1β and IL-8, in response to infection by the ectoparasitic, Chondracanthus goldsmidi. Fish & Shellfish Immunology, 26(5):773-786.
De Jesus E G, Hirano T, Inui Y. 1991. Changes in cortisol and thyroid hormone concentrations during early development and metamorphosis in the Japanese flounder, Paralichthys olivaceus. General and Comparative Endocrinology, 82(3):369-376.
Ellis A E. 2001. Innate host defense mechanisms of fish against viruses and bacteria. Developmental & Comparative Immunology, 25(8-9):827-839.
Forlenza M, Magez S, Scharsack J P, Westphal A, Savelkoul H F J, Wiegertjes G F. 2009. Receptor-mediated and lectinlike activities of carp (Cyprinus carpio) TNF-α. Journal of Immunology, 183(8):5 319-5 332.
García-Castillo J, Chaves-Pozo E, Olivares P, Pelegrín P, Meseguer J, Mulero V. 2004. The tumor necrosis factor α of the bony fish seabream exhibits the in vivo proinflammatory and proliferative activities of its mammalian counterparts, yet it functions in a speciesspecific manner. Cellular and Molecular Life Sciences CMLS, 61(11):1 331-1 340.
García-Castillo J, Pelegrín P, Mulero V, Meseguer J. 2002.Molecular cloning and expression analysis of tumor necrosis factor α from a marine fish reveal its constitutive expression and ubiquitous nature. Immunogenetics, 54(3):200-207.
Grayfer L, Walsh J G, Belosevic M. 2008. Characterization and functional analysis of goldfish (Carassius auratus L.)tumor necrosis factor-alpha. Developmental & Comparative Immunology, 32(5):532-543.
Han J, Brown T, Beutler B. 1990. Endotoxin-responsive sequences control cachectin/tumor necrosis factor biosynthesis at the translational level. Journal of Experimental Medicine, 171(2):465-475.
Hawiger J. 2001. Innate immunity and inflammation:a transcriptional paradigm. Immunologic Research, 23(2-3):99-109.
Hirono I, Nam B H, Kurobe T, Aoki T. 2000. Molecular cloning, characterization, and expression of TNF cDNA and gene from Japanese flounder Paralychthys olivaceus.Journal of Immunology, 165(8):4 423-4 427.
Hong S, Li R G, Xu Q Q, Secombes C J, Wang T H. 2013. Two types of TNF-α Exist in teleost fish:phylogeny, expression, and bioactivity analysis of Type-II TNF-α3 in rainbow trout Oncorhynchus mykiss. Journal of Immunology, 191(12):5 959-5 972.
Hwang S D, Shim S H, Kwon M, Chae Y S, Shim W J, Jung J H, Kim J W, Park C I. 2014. Molecular cloning and expression analysis of two lipopolysaccharide-induced TNF-α factors (LITAFs) from rock bream, Oplegnathus fasciatus. Fish & Shellfish Immunology, 36(2):467-474.
Idriss H T, Naismith J H. 2000. TNFα and the TNF receptor superfamily:structure-function relationship(s).Microscopy Research and Technique, 50(3):184-195.
Inui Y, Miwa S. 1985. Thyroid hormone induces metamorphosis of flounder larvae. General and Comparative Endocrinology, 60(3):450-454.
Jang C H, Choi J H, Byun M S, Jue D M. 2006. Chloroquine inhibits production of TNF-α, IL-1β and IL-6 from lipopolysaccharide-stimulated human monocytes/macrophages by different modes. Rheumatology, 45(6):703-710.
Kalashnikova S A, Goryachev A N, Novochadov V V, Shchyogolev A I. 2009. Thyroid modulation of TNFdependent apoptosis and formation of chronic liver disease in endogenous intoxication in rats. Bulletin of Experimental Biology and Medicine, 147(2):240-244.
Kim B S, Nam B H, Kim J W, Park H J, Song J H, Park C I. 2011. Molecular characterisation and expression analysis of a fish-egg lectin in rock bream, and its response to bacterial or viral infection. Fish & Shellfish Immunology, 31(6):1 201-1 207.
Kiss-Toth E, Harlock E, Lath D, Quertermous T, Wilkinson J M. 2013. A TNF Variant that associates with susceptibility to musculoskeletal disease modulates thyroid hormone receptor binding to control promoter activation. PLoS One, 8(9):e76034.
Klaren P H M, Wunderink Y S, Yúfera M, Mancera J M, Flik G. 2008. The thyroid gland and thyroid hormones in Senegalese sole (Solea senegalensis) during early development and metamorphosis. General and Comparative Endocrinology, 155(3):686-694.
Li H J, Fan J F, Liu S X, Yang Q, Mu G Q, He C B. 2012.Characterization of a myostatin gene (MSTN1) from spotted halibut (Verasper variegatus) and association between its promoter polymorphism and individual growth performance. Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology, 161(4):315-322.
Li H J, Jiang L X, Han J B, Su H, Yang Q, He C B. 2011. Major histocompatibility complex class IIA and IIB genes of the spotted halibut Verasper variegatus:genomic structure, molecular polymorphism, and expression analysis. Fish Physiology and Biochemistry, 37(4):767-780.
Li M F, Zhang J. 2016. CsTNF1, a teleost tumor necrosis factor that promotes antibacterial and antiviral immune defense in a manner that depends on the conserved receptor binding site. Developmental & Comparative Immunology, 55:65-75.
Liu Y W, Chan W K. 2002. Thyroid hormones are important for embryonic to larval transitory phase in zebrafish.Differentiation, 70(1):36-45.
Locksley R M, Killeen N, Lenardo M J. 2001. The TNF and TNF receptor superfamilies:integrating mammalian biology. Cell, 104(4):487-501.
Loetscher H, Steinmetz M, Lesslauer W. 1991. Tumor necrosis factor:receptors and inhibitors. Cancer Cells, 3(6):221-226.
Marchand O, Duffraisse M, Triqueneaux G, Safi R, Laudet V. 2004. Molecular cloning and developmental expression patterns of thyroid hormone receptors and T3 target genes in the turbot (Scophtalmus maximus) during postembryonic development. General and Comparative Endocrinology, 135(3):345-357.
Marchler-Bauer A, Anderson J B, Chitsaz F, Derbyshire M K, DeWeese-Scott C, Fong J H, Geer L Y, Geer R C, Gonzales N R, Gwadz M, He S Q, Hurwitz D I, Jackson J D, Ke Z X, Lanczycki C J, Liebert C A, Liu C L, Lu F, Lu S N, Marchler G H, Mullokandov M, Song J S, Tasneem A, Thanki N, Yamashita R A, Zhang D C, Zhang N G, Bryant S H. 2009. CDD:specific functional annotation with the Conserved Domain Database. Nucleic Acids Research, 37(S1):205-210.
Marchler-Bauer A, Bryant S H. 2004. CD-Search:protein domain annotations on the fly. Nucleic Acids Research, 32(S2):327-331.
Marchler-Bauer A, Derbyshire M K, Gonzales N R, Lu S N, Chitsaz F, Geer L Y, Geer R C, He J N, Gwadz M, Hurwitz D I, Lanczycki C J, Lu F, Marchler G H, Song J S, Thanki N, Wang Z X, Yamashita R A, Zhang D C, Zheng C J, Bryant S H. 2015. CDD:NCBI's conserved domain database. Nucleic Acids Research, 43(D1):D222-D226.
Marchler-Bauer A, Lu S N, Anderson J B, Chitsaz F, Derbyshire M K, DeWeese-Scott C, Fong J H, Geer L Y, Geer R C, Gonzales N R, Gwadz M, Hurwitz D I, Jackson J D, Ke Z X, Lanczycki C J, Lu F, Marchler G H, Mullokandov M, Omelchenko M V, Robertson C L, Song J S, Thanki N, Yamashita R A, Zhang D C, Zhang N Q, Zheng C J, Bryant S H. 2011. CDD:a conserved domain database for the functional annotation of proteins. Nucleic Acids Research, 39(D1):D225-D229.
McGeehan G M, Becherer J D, Bast R C Jr, Boyer C M, Champion B, Connolly K M, Conway J G, Furdon P, Karp S, Kidao S, McElroy A B, Nichols J, Pryzwansky K M, Schoenen F, Sekut L, Truesdale A, Verghese M, Warner J, Ways J P. 1994. Regulation of tumour necrosis factor-α processing by a metalloproteinase inhibitor. Nature, 370(6490):558-561.
Nascimento D S, Pereira P J B, Reis M I R, Do Vale A, Zou J, Silva M T, Secombes C J, Dos Santos N M S. 2007. Molecular cloning and expression analysis of sea bass(Dicentrarchus labrax L.) tumor necrosis factor-α(TNF-α). Fish & Shellfish Immunology, 23(3):701-710.
Olleros M L, Guler R, Corazza N, Vesin D, Eugster H P, Marchal G, Chavarot P, Mueller C, Garcia I. 2002.Transmembrane TNF induces an efficient cell-mediated immunity and resistance to Mycobacterium bovis bacillus Calmette-Guerin infection in the absence of secreted TNF and lymphotoxin-α. Journal of Immunology, 168(7):3 394-3 401.
Ordás M C, Costa M M, Roca F J, López-Castejón G, Mulero V, Meseguer J, Figueras A, Novoa B. 2007. Turbot TNFα gene:molecular characterization and biological activity of the recombinant protein. Molecular Immunology, 44(4):389-400.
Pasparakis M, Alexopoulou L, Episkopou V, Kollias G. 1996.Immune and inflammatory responses in TNF α-deficient mice:a critical requirement for TNF alpha in the formation of primary B cell follicles, follicular dendritic cell networks and germinal centers, and in the maturation of the humoral immune response. Journal of Experimental Medicine, 184(4):1 397-1 411.
Pennica D, Nedwin G E, Hayflick J S, Seeburg P H, Derynck R, Palladino M A, Kohr W J, Aggarwal B B, Goeddel D V. 1984. Human tumour necrosis factor:precursor structure, expression and homology to lymphotoxin. Nature, 312(5996):724-729.
Power D M, Llewellyn L, Faustino M, Nowell M A, Björnsson B T, Einarsdottir I E, Canario A V M, Sweeney G E. 2001.Thyroid hormones in growth and development of fish.Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology, 130(4):447-459.
Praveen K, Evans D L, Jaso-Friedmann L. 2006. Constitutive expression of tumor necrosis factor-alpha in cytotoxic cells of teleosts and its role in regulation of cell-mediated cytotoxicity. Molecular Immunology, 43(3):279-291.
Qi P Z, Xie C X, Guo B Y, Wu C W. 2016. Dissecting the role of transforming growth factor-β1 in topmouth culter immunobiological activity:a fundamental functional analysis. Scientific Reports, 6:27179.
Rink L, Kirchner H. 1996. Recent progress in the tumor necrosis factor-α field. International Archives of Allergy and Immunology, 111(3):199-209.
Roca F J, Mulero I, López-Muñoz A, Sepulcre M P, Renshaw S A, Meseguer J, Mulero V. 2008. Evolution of the inflammatory response in vertebrates:fish TNF-α is a powerful activator of endothelial cells but hardly activates phagocytes. Journal of Immunology, 181(7):5 071-5 081.
Sachs A B. 1993. Messenger RNA degradation in eukaryotes.Cell, 74(3):413-421.
Saeij J P J, Stet R J M, De Vries B J, Van Muiswinkel W B, Wiegertjes G F. 2003. Molecular and functional characterization of carp TNF:a link between TNF polymorphism and trypanotolerance. Developmental and Comparative Immunology, 27(1):29-41.
Saunders B M, Tran S, Ruuls S, Sedgwick J D, Briscoe H, Britton W J. 2005. Transmembrane TNF is sufficient to initiate cell migration and granuloma formation and provide acute, but not long-term, control ofMycobacterium tuberculosis infection. Journal of Immunology, 174(8):4 852-4 859.
Savan R, Sakai M. 2004. Presence of multiple isoforms of TNF alpha in carp (Cyprinus carpio L.):genomic and expression analysis. Fish & Shellfish Immunology, 17(1):87-94.
Sherry B, Cerami A. 1988. Cachectin/tumor necrosis factor exerts endocrine, paracrine, and autocrine control of inflammatory responses. Journal of Cell Biology, 107(4):1 269-1 277.
Singer S J. 1990. The structure and insertion of integral proteins in membranes. Annual Review of Cell Biology, 6:247-296.
Teles M, Mackenzie S, Boltaña S, Callol A, Tort L. 2011. Gene expression and TNF-alpha secretion profile in rainbow trout macrophages following exposures to copper and bacterial lipopolysaccharide. Fish & Shellfish Immunology, 30(1):340-346.
Uysal K T, Wiesbrock S M, Marino M W, Hotamisligil G S. 1997. Protection from obesity-induced insulin resistance in mice lacking TNF-α function. Nature, 389(6651):610-614.
Wang Y S, Bu L Z, Yang L L, Li H Y, Zhang S C. 2016.Identification and functional characterization of fish-egg lectin in zebrafish. Fish & Shellfish Immunology, 52:23-30.
Wride M A, Sanders E J. 1995. Potential roles for tumour necrosis factor α during embryonic development. Anatomy and Embryology, 191(1):1-10.
Xiao J, Zhou Z C, Chen C, Huo W L, Yin Z X, Weng S P, Chan S M, Yu X Q, He J G. 2007. Tumor necrosis factor-alpha gene from mandarin fish, Siniperca chuatsi:Molecular cloning, cytotoxicity analysis and expression profile.Molecular Immunology, 44(14):3 615-3 622.
Xu Y J, Liu X Z, Liao M J, Wang H P, Wang Q Y. 2012.Molecular cloning and differential expression of three GnRH genes during ovarian maturation of spotted halibut, Verasper variegatus. Journal of Experimental Zoology, Part A, Ecological Genetics and Physiology, 317(7):434-446.
Yamano K, Araki K, Sekikawa K, Inui Y. 1994. Cloning of thyroid hormone receptor genes expressed in metamorphosing flounder. Developmental Genetics 15(4):378-382.
Yamano K, Miwa S. 1998. Differential gene expression of thyroid hormone receptor α and β in fish development.General and Comparative Endocrinology, 109(1):75-85.
Zhang A Y, Chen D Y, Wei H, Du L Y, Zhao T Q, Wang X Y, Zhou H. 2012. Functional characterization of TNF-α in grass carp head kidney leukocytes:Induction and involvement in the regulation of NF-κB signaling. Fish & Shellfish Immunology, 33(5):1 123-1 132.
Zhou Z X, Zhang B C, Sun L. 2014. Poly(I:C) induces antiviral immune responses in Japanese flounder (Paralichthys olivaceus) that require TLR3 and MDA5 and is negatively regulated by Myd88. PLoS One, 9(11):e112918.
Zou J, Secombes C J, Long S, Miller N, Clem L W, Chinchar V G. 2003. Molecular identification and expression analysis of tumor necrosis factor in channel catfish(Ictalurus punctatus). Developmental & Comparative Immunology, 27(10):845-858.
Zou J, Wang T, Hirono I, Aoki T, Inagawa H, Honda T, Soma G, Ototake M, Nakanishi T, Ellis A E, Secombes C J. 2002. Differential expression of two tumor necrosis factor genes in rainbow trout, Oncorhynchus mykiss.Developmental & Comparative Immunology, 26(2):161-172.