Molecular Biology Reports

, Volume 37, Issue 3, pp 1523–1529 | Cite as

Molecular cloning of liver Wap65 cDNA in ayu (Plecoglossus altivelis) and mRNA expression changes following Listonella anguillarum infection

  • Yu H. Shi
  • Jiong ChenEmail author
  • Chang H. Li
  • Ming Y. Li


The teleost warm temperature acclimation related 65 kDa protein (Wap65) is a plasma glycoprotein with the potential roles in heat adaptation, heme recycling, immune response and copper metabolism. It is most homologous to the mammalian hemopexin, which is the plasma transporter of heme. A full-length cDNA clone of the Wap65 gene, 1,534 bp in size, was isolated from the fish ayu (Plecoglossus altivelis). Its deduced amino acid sequence of 439 residues had 60.4–65.4% and 38.3–47.3% identical to fish Wap65-2-type and Wap65-1-type sequences, respectively. In phylogenetic analysis, aWap65 grouped tightly with those fish Wap65-2-type sequences. In healthy control fish, the highest mRNA signal for aWap65 was from the liver, moderately high in brain and gill, and but weaker in spleen, kidney, muscle, heart and intestine. In Listonella anguillarum-infected fish, aWap65 transcripts were significantly increased in liver, while no obvious changes in other tissues at 12 hpi. However, aWap65 transcripts were significantly increased in various tissues at 24 hpi when hemolysis developing, suggesting that aWap65 might be involved in the immune response of ayu.


Plecoglossus altivelis Wap65 Sequence analysis mRNA expression profile Bacterial infection 



Warm-temperature-acclimation-associated 65-kDa protein




Ayu Wap65



We thank Dr M. J. Adams, Rothamsted Research, Harpenden, UK for help in correcting the English of the manuscript. The project was supported by the 973 Program (2008CB117015), the S&T Programme of Ningbo Sci-Tech Bureau (2007C10081, 2007A31004), the Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT) (IRT0734) and the KC Wong Magna Fund in Ningbo University.


  1. 1.
    Kikuchi K, Watabe S, Suzuki Y, Aida K, Nakajima H (1993) The 65-kDa cytosolic protein associated with warm temperature acclimation in goldfish, Carassius auratus. J Comp Physiol B 163:349–354. doi: 10.1007/BF00265637 CrossRefGoogle Scholar
  2. 2.
    Aliza D, Ismail IS, Kuah MK, Shu-Chien AC, Muhammad TST (2008) Identification of Wap65, a human homologue of hemopexin as a copper-inducible gene in swordtail fish, Xiphophorus helleri. Fish Physiol Biochem 34:129–138. doi: 10.1007/s10695-007-9153-6 CrossRefPubMedGoogle Scholar
  3. 3.
    Choi CY, An KW, Choi YK, Jo PG, Min BH (2008) Expression of warm temperature acclimation-related protein 65-kDa (Wap65) mRNA, and physiological changes with increasing water temperature in black porgy, Acanthopagrus schlegeli. J Exp Zool Part A Ecol Genet Physiol 309(4):206–214. doi: 10.1002/jez.449 CrossRefGoogle Scholar
  4. 4.
    Hirayama M, Kobiyama A, Kinoshita S, Watabe S (2004) The occurrence of two types of hemopexin-like protein in medaka and differences in their affinity to heme. J Exp Biol 207(8):1387–1398. doi: 10.1242/jeb.00897 CrossRefPubMedGoogle Scholar
  5. 5.
    Hirayama M, Nakaniwa M, Ikeda D, Hirazawa N, Otaka T, Mitsuboshi T, Shirasu K, Watabe S (2003) Primary structures and gene organizations of two types of Wap65 from the pufferfish Takifugu rubripes. Fish Physiol Biochem 29:211–224CrossRefGoogle Scholar
  6. 6.
    Kikuchi K, Yamashita M, Watabe S, Aida K (1995) The warm temperature acclimation-related 65-kDa protein, Wap65, in goldfish and its gene expression. J Biol Chem 270:17087–17092. doi: 10.1074/jbc.270.25.14851 CrossRefPubMedGoogle Scholar
  7. 7.
    Kinoshita S, Itoi S, Watabe S (2001) cDNA cloning and characterization of the warm-temperature-acclimation-associated protein Wap65 from carp Cyprinus carpio. Fish Physiol Biochem 24:125–134. doi: 10.1023/A:1011939321298 CrossRefGoogle Scholar
  8. 8.
    Nakaniwa M, Hirayama M, Shimizu A, Sasaki T, Asakawa S, Shimizu N, Watabe S (2005) Genomic sequences encoding two types of medaka hemopexin-like protein Wap65, and their gene expression profiles in embryos. J Exp Biol 208:1915–1925. doi: 10.1242/jeb.01570 CrossRefPubMedGoogle Scholar
  9. 9.
    Tolosano E, Altruda F (2002) Hemopexin: structure, function, and regulation. DNA Cell Biol 21(4):297–306. doi: 10.1089/104454902753759717 CrossRefPubMedGoogle Scholar
  10. 10.
    Delanghe JR, Langlois MR (2001) Hemopexin: a review of biological aspects and the role in laboratory medicine. Clin Chim Acta 12(1–2):13–23. doi: 10.1016/S0009-8981(01)00586-1 CrossRefGoogle Scholar
  11. 11.
    Kikuchi K, Watabe S, Aida K (1997) The Wap65 gene expression of goldfish (Carassius auratus) in association with warm water temperature as well as bacterial lipopolysaccharide (LPS). Fish Physiol Biochem 17:423–432. doi: 10.1023/A:1007768531655 CrossRefGoogle Scholar
  12. 12.
    Kikuchi K, Watabe S, Aida K (1998) Isolation of a 65-kDa protein from white muscle of warm temperature-acclimated goldfish (Carassius auratus). Comp Biochem Physiol B Biochem Mol Biol 120:385–391. doi: 10.1016/S0305-0491(98)10045-7 CrossRefPubMedGoogle Scholar
  13. 13.
    Watabe S, Kikuchi K, Aida K (1993) Cold- and warm-temperature acclimation induces specific cytosolic protein in goldfish and carp. Nippon Suisan Gakkai 59:151–156Google Scholar
  14. 14.
    Pickett CL, Auffenberg T, Pesci EC, Sheen VL, Jusuf SS (1992) Iron acquisition and emolysin production by Campylobacter jejuni. Infect Immun 60:3872–3877PubMedGoogle Scholar
  15. 15.
    Barclay R (1985) The role of iron in infection. Med Lab Sci 42:166–177PubMedGoogle Scholar
  16. 16.
    Peatman E, Terhune J, Baoprasertkul P, Xu P, Nandi S, Wang S, Somridhivej B, Kucuktas H, Li P, Dunham R, Liu Z (2008) Microarray analysis of gene expression in the blue catfish liver reveals early activation of the MHC class I pathway after infection with Edwardsiella ictaluri. Mol Immunol 45(2):553–566. doi: 10.1016/j.molimm.2007.05.012 CrossRefPubMedGoogle Scholar
  17. 17.
    Sha Z, Xu P, Takano T, Liu H, Terhune J, Liu Z (2008) The warm temperature acclimation protein Wap65 as an immune response gene: its duplicates are differentially regulated by temperature and bacterial infections. Mol Immunol 45(5):1458–1469. doi: 10.1016/j.molimm.2007.08.012 CrossRefPubMedGoogle Scholar
  18. 18.
    Chen J, Shi YH, Li MY (2008) Molecular cloning of liver angiotensinogen gene in ayu (Plecoglossus altivelis) and mRNA expression changes upon Aeromonas hydrophila infection. Fish Shellfish Immunol 24(5):659–662. doi: 10.1016/j.fsi.2008.01.015 CrossRefPubMedGoogle Scholar
  19. 19.
    Kondo M, Kawai K, Kurohara K, Oshima S (2002) Adherence of Flavobacterium psychrophilum on the body surface of the ayu Plecoglossus altivelis. Microbes Infect 4:279–283. doi: 10.1016/S1286-4579(02)01539-3 CrossRefPubMedGoogle Scholar
  20. 20.
    Nishimori E, Kita-Tsukamoto K, Wakabayashi H (2000) Pseudomonas plecoglossicida sp. nov., the causative agent of bacterial haemorrhagic ascites of ayu, Plecoglossus altivelis. Int J Syst Evol Microbiol 50:83–89PubMedGoogle Scholar
  21. 21.
    Wang WS, Wang DH, Lee JS (1996) An outbreak of Aeromonas salmonicida infection of ayu (Plecoglossus altivelis) in Taiwan. Asian Fish Sci 9:235–238Google Scholar
  22. 22.
    Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599. doi: 10.1093/molbev/msm092 CrossRefPubMedGoogle Scholar
  23. 23.
    Paoli M, Anderson BF, Baker HM, Morgan WT, Smith A, Baker EN (1999) Crystal structure of hemopexin reveals a novel high-affinity heme site formed between two β propeller domains. Nat Struct Biol 6:926–931. doi: 10.1038/13294 CrossRefPubMedGoogle Scholar
  24. 24.
    Ohashi H, Umeda N, Hirazawa N, Ozaki Y, Miura C, Miura T (2007) Purification and identification of a glycoprotein that induces the attachment of oncomiracidia of Neobenedenia girellae (Monogenea, Capsalidae). Int J Parasitol 37(13):1483–1490. doi: 10.1016/j.ijpara.2007.04.024 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Yu H. Shi
    • 1
  • Jiong Chen
    • 1
    Email author
  • Chang H. Li
    • 1
  • Ming Y. Li
    • 1
  1. 1.Key Laboratory of Applied Marine Biotechnology, Ministry of EducationNingbo UniversityNingboPeople’s Republic of China

Personalised recommendations