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Molecular cloning, expression analysis, and the immune-related role of a thymosin β in the goldfish, Carassius auratus

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Abstract

β-Thymosins play critical roles in the regulation of many important physiological processes, but their function in teleost fishes remains poorly understood. In this study, the full-length cDNA coding for a thymosin β (Tβ) was cloned and identified in goldfish, Carassius auratus (gfTβ). The gfTβ cDNA consisted of 653 bp with an open reading frame of 135 bp that encodes a 44 amino acid polypeptide. Sequence analysis revealed one thymosin domain and a highly conserved actin-binding motif (18LKKTET23). Expression of gfTβ transcript was detected ubiquitously in all tissues examined, with relatively higher levels in the brain, intestine, spleen, gill, skin, kidney, and testis. Cadmium and H2O2 exposure induced increases in gfTβ transcript levels in the liver and spleen. Moreover, gfTβ transcription was upregulated in response to LPS challenge in the spleen while Poly I:C treatment did not affect gfTβ expression. In vivo injection of recombinant gfTβ generated from an Escherichia coli system induced expression of T lymphocyte-related genes (RAG1 and CD8α). These results suggest that gfTβ may be involved in the immune response of teleost fishes via modulation of T lymphocyte development.

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References

  • Alexopoulou L, Holt AC, Medzhitov R, Flavell RA (2001) Recognition of double-stranded RNA and activation of NF-kappa B by Toll-like receptor 3. Nature 413(6857):732–738

    Article  CAS  Google Scholar 

  • Anathy V, Kirankumar S, Pandian TJ (2003) Enhanced expression of beta-thymosin mRNA in the ovary of GnRH analog or estradiol-17beta-treated paradise fish, Macropodus opercularis. Comp Biochem Physiol A Mol Integr Physiol 134(3):639–645

    Article  CAS  Google Scholar 

  • Boquet I, Boujemaa R, Carlier MF, Preat T (2000) Ciboulot regulates actin assembly during Drosophila brain metamorphosis. Cell 102(6):797–808

    Article  CAS  Google Scholar 

  • Cesta MF (2006) Normal structure, function, and histology of the spleen. Toxicol Pathol 34(5):455–465

    Article  Google Scholar 

  • Chen W, Zhang Z, Yan F, Jiang X, Qin S, Dong H (2017) Identification of three selenoprotein T paralogs in goldfish (Carassius auratus) and expression analysis in response to environmental stressors. Comp Biochem Physiol B Biochem Mol Biol 203:65–75

    Article  CAS  Google Scholar 

  • dos Remedios CG, Chhabra D, Kekic M, Dedova IV, Tsubakihara M, Berry DA, Nosworthy NJ (2003) Actin binding proteins: regulation of cytoskeletal microfilaments. Physiol Rev 83(2):433–473

    Article  Google Scholar 

  • Erickson-Viitanen S, Horecker BL (1984) Thymosin beta 11: a peptide from trout liver homologous to thymosin beta 4. Arch Biochem Biophys 233(2):815–820

    Article  CAS  Google Scholar 

  • Gan Z, Wang B, Zhou W, Lu Y, Zhu W, Tang J, Jian J, Wu Z (2015) Molecular and functional characterization of CD59 from Nile tilapia (Oreochromis niloticus) involved in the immune response to Streptococcus agalactiae. Fish Shellfish Immunol 44(1):50–59

    Article  CAS  Google Scholar 

  • Gan Z, Wang B, Tang J, Lu Y, Jian J, Wu Z, Nie P (2016) Molecular characterization and expression of CD2 in Nile tilapia (Oreochromis niloticus) in response to Streptococcus agalactiae stimulus. Fish Shellfish Immunol 50:101–108

    Article  CAS  Google Scholar 

  • Goodall GJ, Dominguez F, Horecker BL (1986) Molecular cloning of cDNA for human prothymosin alpha. Proc Natl Acad Sci U S A 83(23):8926–8928

    Article  CAS  Google Scholar 

  • Hall AK (1991) Differential expression of thymosin genes in human tumors and in the developing human kidney. Int J Cancer 48(5):672–677

    Article  CAS  Google Scholar 

  • Hannappel E, Huff T (2003) The thymosins. Prothymosin alpha, parathymosin, and beta-thymosins: structure and function. Vitam Horm 66:257–296

    Article  CAS  Google Scholar 

  • Hermesz E, Abrahám M, Nemcsók J (2001) Tissue-specific expression of two metallothionein genes in common carp during cadmium exposure and temperature shock. Comp Biochem Physiol C Toxicol Pharmacol 128(3):457–465

    Article  CAS  Google Scholar 

  • Huff T, Muller CS, Otto AM, Netzker R, Hannappel E (2001) beta-Thymosins, small acidic peptides with multiple functions. Int J Biochem Cell Biol 33(3):205–220

    Article  CAS  Google Scholar 

  • Kasthuri SR, Premachandra HK, Umasuthan N, Whang I, Lee J (2013) Structural characterization and expression analysis of a beta-thymosin homologue (Tβ) in disk abalone, Haliotis discus discus. Gene 527(1):376–383

    Article  CAS  Google Scholar 

  • Kim JH, Wang SY, Kim IC, Ki JS, Raisuddin S, Lee JS, Han KN (2008) Cloning of a river pufferfish (Takifugu obscurus) metallothionein cDNA and study of its induction profile in cadmium-exposed fish. Chemosphere 71(7):1251–1259

    Article  CAS  Google Scholar 

  • Kumar S, Gupta S (2011) Thymosin beta 4 prevents oxidative stress by targeting antioxidant and anti-apoptotic genes in cardiac fibroblasts. PLoS One 6(10):e26912

    Article  CAS  Google Scholar 

  • Kuzan A (2016) Thymosin β as an actin-binding protein with a variety of functions. Adv Clin Exp Med 25(6):1331–1336

    Article  Google Scholar 

  • Lee HR, Yoon SY, Kang HB, Park S, Kim KE, Cho YH, Kim S, Kim CW, Cho BJ, Lee WJ, Bang SI, Park H, Cho D (2009) Thymosin beta 4 enhances NK cell cytotoxicity mediated by ICAM-1. Immunol Lett 123(1):72–76

    Article  CAS  Google Scholar 

  • Li F, Xiang J (2013) Signaling pathways regulating innate immune responses in shrimp. Fish Shellfish Immunol 34(4):973–980

    Article  CAS  Google Scholar 

  • Li X, Zheng L, Peng F, Qi C, Zhang X, Zhou A, Liu Z, Wu S (2007) Recombinant thymosin beta 4 can promote full-thickness cutaneous wound healing. Protein Expr Purif 56(2):229–236

    Article  CAS  Google Scholar 

  • Li X, Jiang ZX, Hao P, Tang X (2013) Protective effect of thymosin β4 against oxidative damage in cultured rabbit corneal epithelial cells. Zhonghua Yan Ke Za Zhi 49(8):716–722 (in Chinese)

    CAS  PubMed  Google Scholar 

  • Li J, Zhang Y, Liu Y, Zhang Y, Xiang Z, Qu F, Yu Z (2016) A thymosin beta-4 is involved in production of hemocytes and immune defense of Hong Kong oyster, Crassostrea hongkongensis. Dev Comp Immunol 57:1–9

    Article  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4):402–408

    Article  CAS  Google Scholar 

  • Low TL, Hu SK, Goldstein AL (1981) Complete amino acid sequence of bovine thymosin beta 4: a thymic hormone that induces terminal deoxynucleotidyl transferase activity in thymocyte populations. Proc Natl Acad Sci U S A 78(2):1162–1166

    Article  CAS  Google Scholar 

  • Low TL, Liu DT, Jou JH (1992) Primary structure of thymosin beta 12, a new member of the beta-thymosin family isolated from perch liver. Arch Biochem Biophys 293(1):32–39

    Article  CAS  Google Scholar 

  • Malinda KM, Goldstein AL, Kleinman HK (1997) Thymosin beta 4 stimulates directional migration of human umbilical vein endothelial cells. FASEB J 11(6):474–481

    Article  CAS  Google Scholar 

  • Mebius RE, Kraal G (2005) Structure and function of the spleen. Nat Rev Immunol 5(8):606–616

    Article  CAS  Google Scholar 

  • Nam BH, Seo JK, Lee MJ, Kim YO, Kim DG, An CM, Park NG (2015) Functional analysis of Pacific oyster (Crassostrea gigas) β-thymosin: focus on antimicrobial activity. Fish Shellfish Immunol 45(1):167–174

    Article  CAS  Google Scholar 

  • Petrie-Hanson L, Hohn C, Hanson L (2009) Characterization of rag1 mutant zebrafish leukocytes. BMC Immunol 10:8

    Article  Google Scholar 

  • Priyadarshini M, Orosco LA, Panula PJ (2013) Oxidative stress and regulation of Pink1 in zebrafish (Danio rerio). PLoS One 8(11):e81851

    Article  Google Scholar 

  • Qiang J, Tao YF, He J, Xu P, Bao JW, Sun YL (2017) miR-122 promotes hepatic antioxidant defense of genetically improved farmed tilapia (GIFT, Oreochromis niloticus) exposed to cadmium by directly targeting a metallothionein gene. Aquat Toxicol 182:39–48

    Article  CAS  Google Scholar 

  • Rebar RW, Miyake A, Low TL, Goldstein AL (1981) Thymosin stimulates secretion of luteinizing hormone-releasing factor. Science 214(4521):669–671

    Article  CAS  Google Scholar 

  • Shi XZ, Shi LJ, Zhao YR, Zhao XF, Wang JX (2015) β-Thymosins participate in antiviral immunity of red swamp crayfish (Procambarus clarkii). Dev Comp Immunol 51(2):213–225

    Article  CAS  Google Scholar 

  • Stoeva S, Hörger S, Voelter W (1997) A novel β-thymosin from the sea urchin: extending the phylogenetic distribution of β thymosins from mammals to echinoderms. J Pept Sci 3(4):282–290

    Article  CAS  Google Scholar 

  • Suetake H, Araki K, Suzuki Y (2004) Cloning, expression, and characterization of fugu CD4, the first ectothermic animal CD4. Immunogenetics 56(5):368–374

    Article  CAS  Google Scholar 

  • Sun Y, Chen X, Xu Y, Liu Q, Jiang X, Wang S, Guo W, Zhou Y (2017) Thymosin β4 is involved in the antimicrobial immune response of Golden pompano, Trachinotus ovatus. Fish Shellfish Immunol 69:90–98

    Article  CAS  Google Scholar 

  • Takeuchi O, Hoshino K, Kawai T, Sanjo H, Takada H, Ogawa T, Takeda K, Akira S (1999) Differential roles of TLR2 and TLR4 in recognition of gram-negative and gram-positive bacterial cell wall components. Immunity 11(4):443–451

    Article  CAS  Google Scholar 

  • Tang YQ, Yeaman MR, Selsted ME (2002) Antimicrobial peptides from human platelets. Infect Immun 70(12):6524–6533

    Article  CAS  Google Scholar 

  • Wang T, Secombes CJ (2008) Rainbow trout suppressor of cytokine signaling (SOCS)-1, 2 and 3: molecular identification, expression and modulation. Mol Immunol 45(5):1449–1457

    Article  CAS  Google Scholar 

  • Wang Q, Ju X, Chen Y, Dong X, Luo S, Liu H, Zhang D (2016a) Effects of L-carnitine against H2O2-induced oxidative stress in grass carp ovary cells (Ctenopharyngodon idellus). Fish Physiol Biochem 42(3):845–857

    Article  CAS  Google Scholar 

  • Wang M, Zhao X, Kong X, Wang L, Jiao D, Zhang H (2016b) Molecular characterization and expressing analysis of the c-type and g-type lysozymes in Qihe crucian carp Carassius auratus. Fish Shellfish Immunol 52:210–220

    Article  CAS  Google Scholar 

  • Wei C, Kumar S, Kim IK, Gupta S (2012) Thymosin beta 4 protects cardiomyocytes from oxidative stress by targeting anti-oxidative enzymes and anti-apoptotic genes. PLoS One 7(8):e42586

    Article  CAS  Google Scholar 

  • Wu L, Wu X (2009) Molecular cloning and expression analysis of a beta-thymosin homologue from a gastropod abalone, Haliotis diversicolor supertexta. Fish Shellfish Immunol 27(2):379–382

    Article  CAS  Google Scholar 

  • Wu SM, Weng CF, Yu MJ, Lin CC, Chen ST, Hwang JC, Hwang PP (1999) Cadmium-inducible metallothionein in tilapia (Oreochromis mossambicus). Bull Environ Contam Toxicol 62(6):758–768

    Article  CAS  Google Scholar 

  • Xiao Z, Shen J, Feng H, Liu H, Wang Y, Huang R, Guo Q (2015) Characterization of two thymosins as immune-related genes in common carp (Cyprinus carpio L.). Dev Comp Immunol 50(1):29–37

    Article  CAS  Google Scholar 

  • Yialouris PP, Coles B, Tsitsiloni O, Schmid B, Howell S, Aitken A, Voelter W, Haritos AA (1992) The complete sequences of trout (Salmo gairdneri) thymosin beta 11 and its homologue thymosin beta 12. Biochem J 283(Pt2):385–389

    Article  CAS  Google Scholar 

  • Zapata A, Diez B, Cejalvo T, Gutiérrez-de Frías C, Cortés A (2006) Ontogeny of the immune system of fish. Fish Shellfish Immunol 20(2):126–136

    Article  CAS  Google Scholar 

  • Zhang Y, Fu D, Yu F, Liu Q, Yu Z (2011) Two catalase homologs are involved in host protection against bacterial infection and oxidative stress in Crassostrea hongkongensis. Fish Shellfish Immunol 31(6):894–903

    Article  CAS  Google Scholar 

  • Zhu B, Wu ZF, Li J, Wang GX (2011) Single and joint action toxicity of heavy metals on early developmental stages of Chinese rare minnow (Gobiocypris rarus). Ecotoxicol Environ Saf 74(8):2193–2202

    Article  CAS  Google Scholar 

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Acknowledgments

This work is financially supported by the National Natural Science Foundation of China (No. 31101878), the Excellent Youth Foundation of Henan Polytechnic University (No. J2014-02), and the Key Science Research Project in University of Henan Province (No. 16A240001).

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Authors and Affiliations

  1. Department of Biology, Institute of Resources and Environment, Henan Polytechnic University, Jiaozuo, 454000, Henan, China

    Wenbo Chen, Fangfang Yan & Shaozong Qin

  2. Department of Medicine, Huzhou University, Huzhou, 313000, Zhejiang, China

    Haiyan Dong

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  1. Wenbo Chen
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Correspondence to Wenbo Chen or Haiyan Dong.

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Chen, W., Yan, F., Qin, S. et al. Molecular cloning, expression analysis, and the immune-related role of a thymosin β in the goldfish, Carassius auratus. Fish Physiol Biochem 45, 427–437 (2019). https://doi.org/10.1007/s10695-018-0574-1

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