Advertisement

Fish Physiology and Biochemistry

, Volume 45, Issue 1, pp 427–437 | Cite as

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

  • Wenbo ChenEmail author
  • Fangfang Yan
  • Shaozong Qin
  • Haiyan DongEmail author
Article

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.

Keywords

Thymosin β Cloning Immune response Cadmium Oxidative stress Carassius auratus 

Notes

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).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

10695_2018_574_MOESM1_ESM.docx (12 kb)
ESM 1 (DOCX 24.1 kb)
10695_2018_574_MOESM2_ESM.docx (12 kb)
ESM 2 (DOCX 24.1 kb)

References

  1. 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):732–738Google Scholar
  2. 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):639–645Google Scholar
  3. Boquet I, Boujemaa R, Carlier MF, Preat T (2000) Ciboulot regulates actin assembly during Drosophila brain metamorphosis. Cell 102(6):797–808Google Scholar
  4. Cesta MF (2006) Normal structure, function, and histology of the spleen. Toxicol Pathol 34(5):455–465Google Scholar
  5. 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 203:65–75Google Scholar
  6. 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):433–473Google Scholar
  7. Erickson-Viitanen S, Horecker BL (1984) Thymosin beta 11: a peptide from trout liver homologous to thymosin beta 4. Arch Biochem Biophys 233(2):815–820Google Scholar
  8. 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):50–59Google Scholar
  9. 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 50:101–108Google Scholar
  10. Goodall GJ, Dominguez F, Horecker BL (1986) Molecular cloning of cDNA for human prothymosin alpha. Proc Natl Acad Sci U S A 83(23):8926–8928Google Scholar
  11. Hall AK (1991) Differential expression of thymosin genes in human tumors and in the developing human kidney. Int J Cancer 48(5):672–677Google Scholar
  12. Hannappel E, Huff T (2003) The thymosins. Prothymosin alpha, parathymosin, and beta-thymosins: structure and function. Vitam Horm 66:257–296Google Scholar
  13. 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):457–465Google Scholar
  14. 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):205–220Google Scholar
  15. 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):376–383Google Scholar
  16. 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):1251–1259Google Scholar
  17. 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):e26912Google Scholar
  18. Kuzan A (2016) Thymosin β as an actin-binding protein with a variety of functions. Adv Clin Exp Med 25(6):1331–1336Google Scholar
  19. 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):72–76Google Scholar
  20. Li F, Xiang J (2013) Signaling pathways regulating innate immune responses in shrimp. Fish Shellfish Immunol 34(4):973–980Google Scholar
  21. 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):229–236Google Scholar
  22. 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) Google Scholar
  23. 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 57:1–9Google Scholar
  24. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4):402–408Google Scholar
  25. 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):1162–1166Google Scholar
  26. 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):32–39Google Scholar
  27. Malinda KM, Goldstein AL, Kleinman HK (1997) Thymosin beta 4 stimulates directional migration of human umbilical vein endothelial cells. FASEB J 11(6):474–481Google Scholar
  28. Mebius RE, Kraal G (2005) Structure and function of the spleen. Nat Rev Immunol 5(8):606–616Google Scholar
  29. 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):167–174Google Scholar
  30. Petrie-Hanson L, Hohn C, Hanson L (2009) Characterization of rag1 mutant zebrafish leukocytes. BMC Immunol 10:8Google Scholar
  31. Priyadarshini M, Orosco LA, Panula PJ (2013) Oxidative stress and regulation of Pink1 in zebrafish (Danio rerio). PLoS One 8(11):e81851Google Scholar
  32. 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 182:39–48Google Scholar
  33. Rebar RW, Miyake A, Low TL, Goldstein AL (1981) Thymosin stimulates secretion of luteinizing hormone-releasing factor. Science 214(4521):669–671Google Scholar
  34. 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):213–225Google Scholar
  35. 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):282–290Google Scholar
  36. Suetake H, Araki K, Suzuki Y (2004) Cloning, expression, and characterization of fugu CD4, the first ectothermic animal CD4. Immunogenetics 56(5):368–374Google Scholar
  37. 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 69:90–98Google Scholar
  38. 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):443–451Google Scholar
  39. Tang YQ, Yeaman MR, Selsted ME (2002) Antimicrobial peptides from human platelets. Infect Immun 70(12):6524–6533Google Scholar
  40. 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):1449–1457Google Scholar
  41. 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):845–857Google Scholar
  42. 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 52:210–220Google Scholar
  43. 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):e42586Google Scholar
  44. 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):379–382Google Scholar
  45. 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):758–768Google Scholar
  46. 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):29–37Google Scholar
  47. 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):385–389Google Scholar
  48. 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):126–136Google Scholar
  49. 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):894–903Google Scholar
  50. 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):2193–2202Google Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of Biology, Institute of Resources and EnvironmentHenan Polytechnic UniversityJiaozuoChina
  2. 2.Department of MedicineHuzhou UniversityHuzhouChina

Personalised recommendations