, Volume 31, Issue 4, pp 527–537 | Cite as

Six indicator genes for zinc (Zn) homeostasis in freshwater teleost yellow catfish Pelteobagrus fulvidraco: molecular characterization, mRNA tissue expression and transcriptional changes to Zn exposure

  • Guang-Hui Chen
  • Zhi Luo
  • Chuan-Chuan Wei
  • Dan-Dan Li
  • Ya-Xiong Pan


Excessive Zn in the aquatic environment can be toxic and causes dysfunction in Zn homeostasis for fish, which ultimately influences the function of various biological processes. Zn homeostasis is controlled by Zn transporters. This study cloned and characterized the full-length cDNA sequences of six Zn transport-relevant genes (ZnT1, ZnT5, ZnT7, ZIP4, ZIP5 and MTF-1) from yellow catfish Pelteobagrus fulvidraco. The six genes share similar domains to their corresponding members of mammals. Their mRNA amounts were widely existent across eight tissues (intestine, liver, brain, heart, gill, muscle, spleen and mesenteric fat), but relatively predominant in the liver and intestine. On day 28, Zn exposure tended to increase transcript levels of ZnT1, ZnT5 and MTF-1, decrease hepatic ZIP5 expression, but did not significantly affect the expression of ZnT7 and ZIP4. On day 56, Zn exposure tended to increase transcript levels of ZnT1 and MTF-1, down-regulate hepatic mRNA amounts of ZIP4 and ZIP5; among three Zn treatments, ZnT5 expression in the 0.5 mg Zn/L group and ZnT7 expression in the 0.25 mg Zn/L group were the highest. The mRNA abundances of these genes showed Zn concentration- and exposure time-dependent manners. For the first time, we characterized the full-length cDNA sequences of six Zn transport-relevant genes in fish, explored their tissue expression profiles and transcriptional responses to Zn exposure. Our study built good basis for further investigating their physiological functions of these genes and provided new insights into the regulatory mechanisms of Zn homeostasis in fish.


Pelteobagrus fulvidraco Zn exposure Zn transporter Zn homeostasis 



Amino acid


One-way analysis of variance


Aspartic acid




Metal response element binding transcription factor-1


Nuclear exclusion sequence


Nuclear localization sequence


Open reading frame


Pelteobagrus fulvidraco


Real-time fluorescence quantitative PCR


Standard error of means


Signal peptide


Transmembrane domain





This work was supported by the National Natural Science Foundation of China (Grant no. 31422056).

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest with the contents of this article.

Supplementary material

10534_2018_99_MOESM1_ESM.doc (733 kb)
Supplementary material 1 (DOC 733 kb)


  1. Andrews GK (2001) Cellular zinc sensors: MTF-1 regulation of gene expression. Biometals 14:223–237CrossRefPubMedGoogle Scholar
  2. Balesaria S, Hogstrand C (2006) Identification, cloning and characterization of a plasma membrane zinc efflux transporter, TrZnT-1, from fugu pufferfish (Takifugu rubripes). Biochem J 394:485–493CrossRefPubMedPubMedCentralGoogle Scholar
  3. Chen WY, John JAC, Lin CH, Chang CY (2002) Molecular cloning and developmental expression of zinc finger transcription factor MTF-1 gene in zebrafish, Danio rerio. Biochem Biophys Res Commun 29:798–805CrossRefGoogle Scholar
  4. Chen F, Luo Z, Fan YF, Wu K, Pan YX, Liu X, Zhang LH, Song YF (2016) Five metal elements homeostasis-related genes in Synechogobius hasta: molecular characterization, tissue expression and transcriptional response to Cu and Fe exposure. Chemosphere 159:392–402CrossRefPubMedGoogle Scholar
  5. Chen GH, Luo Z, Chen F, Shi X, Song YF, You WJ, Liu X (2017) PPARα, PPARγ and SREBP-1 pathways mediated waterborne iron (Fe)-induced reduction in hepatic lipid deposition of javelin goby Synechogobius hasta. Comp Biochem Physiol 197C:8–18Google Scholar
  6. Cousins RJ (1985) Absorption, transport, and hepatic metabolism of copper and zinc: special reference to metallothionein and ceruloplasmin. Physiol Rev 65:238–309CrossRefPubMedGoogle Scholar
  7. Cragg RA, Christie GR, Phillips SR, Russi RM, Kury S, Mathers JC, Taylor PM, Ford D (2002) A novel zinc-regualted human zinc transporter, hZTL1, is localized to the enterocyte apical membrane. J Biol Chem 277:22789–22797CrossRefPubMedGoogle Scholar
  8. Cragg RA, Phillips SR, Piper JM, Varma JS, Campbell FC, Mathers JC, Ford D (2005) Homeostatic regulation of zinc transporters in the human small intestine by dietary zinc supplementation. Gut 54:469–478CrossRefPubMedPubMedCentralGoogle Scholar
  9. Dufner-Beattie J, Wang F, Kuo YM, Gitschier J, Eide D, Andrews GK (2003) The acrodermatitis enteropathica gene ZIP4 encodes a tissue-specific, zinc-regulated zinc transporter in mice. J Biol Chem 278:33474–33481CrossRefPubMedGoogle Scholar
  10. Dufner-Beattie J, Kuo YM, Gitschier J, Andrews GK (2004) The adaptive response to dietary zinc in mice involves the differential cellular localization and zinc regulation of the zinc transporters ZIP4 and ZIP5. J Biol Chem 279:49082–49090CrossRefPubMedGoogle Scholar
  11. Feeney GP, Zheng D, Kille P, Hogstrand C (2005) The phylogeny of teleost ZIP and ZnT zinc transporters and their tissue specific expression and response to zinc in zebrafish. Biochim Biophys Acta 1732:88–95CrossRefPubMedGoogle Scholar
  12. Gaither LA, Eide DJ (2000) Functional expression of the human hZIP2 zinc transporter. J Biol Chem 275:5560–5564CrossRefPubMedGoogle Scholar
  13. Gaither LA, Eide DJ (2001) Eukaryotic zinc transporters and their regulation. Biometals 14:251–270CrossRefPubMedGoogle Scholar
  14. Geiser J, De Lisle RC, Andrews GK (2013) The zinc transporter Zip5 (Slc39a5) regulates intestinal zinc excretion and protects the pancreas against zinc toxicity. PLoS ONE 8:e82149CrossRefPubMedPubMedCentralGoogle Scholar
  15. Guerrerio AL, Berg JM (2004) Metal ion affinities of the zinc finger domains of the metal responsive element-binding transcription factor-1 (MTF1). Biochemistry 43:5437–5444CrossRefPubMedGoogle Scholar
  16. Heath AG (1987) Water pollution and fish physiology. CRC Press, Boca Raton, p 245Google Scholar
  17. Ho E, Dukovcic S, Hobson B, Wong CP, Miller G, Hardin K, Tanguay RL (2012) Zinc transporter expression in zebrafish (Danio rerio) during development. Comp Biochem Physiol 155C:26–32Google Scholar
  18. Hoch E, Lin W, Chai J, Hershfinkel M, Fu D, Sekler I (2012) Histidine pairing at the metal transport site of mammalian ZnT transporters controls Zn2+ over Cd2+ selectivity. Proc Natl Acad Sci USA 109:7202–7207CrossRefPubMedGoogle Scholar
  19. Huang L, Tepaamorndech S (2013) The SLC30 family of zinc transporters: a review of current understanding of their biological and pathophysiological roles. Mol Aspects Med 34:548–560CrossRefPubMedGoogle Scholar
  20. Ishihara K, Yamazaki T, Ishida Y, Suzuki T, Oda K, Nagao M, Yamaguchi-Iwai Y, Kambe T (2006) Zinc transport complexes contribute to the homeostatic maintenance of secretory pathway function in vertebrate cells. J Biol Chem 281:17743–17750CrossRefPubMedGoogle Scholar
  21. Jones DT, Taylor WR, Thornton JM (1992) The rapid generation of mutation data matrices from protein sequences. Comput Appl Biosci 8:275–282PubMedGoogle Scholar
  22. Kambe T, Narita H, Yamaguchi-Iwai Y, Hirose J, Amano T, Sugiura N (2002) Cloning and characterization of a novel mammalian zinc transporter, zinc transporter 5, abundantly expressed in pancreatic beta cells. J Biol Chem 277:19049–19055CrossRefPubMedGoogle Scholar
  23. Kambe T, Yamaguchiiwai Y, Sasaki R, Nagao M (2004) Overview of mammalian zinc transporters. Cell Mol Life Sci 61:49–68CrossRefPubMedGoogle Scholar
  24. Katoh K, Misawa K, Kuma KI, Miyata T (2002) MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res 30:3059–3066CrossRefPubMedPubMedCentralGoogle Scholar
  25. Kim AH, Sheline CT, Tian M, Higashi T, McMahon RJ, Cousins RJ (2000) L-type Ca2+ channel-mediated Zn2+ toxicity and modulation by ZnT-1 in PC12 cells. Brain Res 886:99–107CrossRefPubMedGoogle Scholar
  26. Kirschke CP, Huang L (2003) ZnT7, a novel mammalian zinc transporter, accumulates zinc in the Golgi apparatus. J Biol Chem 278:4096–4102CrossRefPubMedGoogle Scholar
  27. Laity JH, Andrews GK (2007) Understanding the mechanisms of zinc-sensing by metal-response element binding transcription factor-1 (MTF-1). Arch Biochem Biophys 463:201–210CrossRefPubMedGoogle Scholar
  28. Langmade SJ, Ravindra R, Daniels PJ, Andrews GK (2000) The transcription factor MTF-1 mediates metal regulation of the mouse ZnT1 gene. J Biol Chem 275:34803–34809CrossRefPubMedGoogle Scholar
  29. Liuzzi JP, Cousins RJ (2004) Mammalian zinc transporters. Annu Rev Nutr 24:151–172CrossRefPubMedGoogle Scholar
  30. Liuzzi JP, Blanchard RK, Cousins RJ (2001) Differential regulation of zinc transporter 1, 2 and 4 mRNA expression by dietary zinc in rats. J Nutr 131:46–52CrossRefPubMedGoogle Scholar
  31. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using realtime quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  32. Mao X, Kim BE, Wang F, Eide DJ, Petris MJ (2007) A histidine-rich cluster mediates the ubiquitination and degradation of the human zinc transporter, hZIP4, and protects against zinc cytotoxicity. J Biol Chem 282:6992–7000CrossRefPubMedGoogle Scholar
  33. McMahon RJ, Cousins RJ (1998) Regulation of the zinc transporter ZnT-1 by dietary zinc. Proc Natl Acad Sci USA 95:4841–4846CrossRefPubMedGoogle Scholar
  34. Muylle F, Robbens J, De Coen W, Timmermans JP, Blust R (2006) Cadmium and zinc induction of ZnT-1 mRNA in an established carp cell line. Comp Biochem Physiol 143C:242–251Google Scholar
  35. Palmiter RD, Findley SD (1995) Cloning and functional characterization of a mammalian zinc transporter that confers resistance to zinc. EMBO J 14:639–649PubMedPubMedCentralGoogle Scholar
  36. Penn O, Privman E, Ashkenazy H, Landan G, Graur D, Pupko T (2010) Guidance: a web server for assessing alignment confidence scores. Nucleic Acids Res 38:W23–W28CrossRefPubMedPubMedCentralGoogle Scholar
  37. Potter BM, Feng LS, Parasuram P, Matskevich VA, Wilson JA, Andrews GK, Laity JH (2005) The six zinc fingers of metal-responsive element binding transcription factor-1 form stable and quasi-ordered structures with relatively small differences in zinc affinities. J Biol Chem 280:28529–28540CrossRefPubMedGoogle Scholar
  38. Radtke F, Heuchel R, Georgiev O, Hergersberg M, Gariglio M, Dembic Z, Schaffner W (1993) Cloned transcription factor MTF-1 activates the mouse metallothionein I promoter. EMBO J 12:1355–1362PubMedPubMedCentralGoogle Scholar
  39. Saydam N, Georgiev O, Nakano MY, Greber UF, Schaffner W (2001) Nucleo-cytoplasmic trafficking of metal-regulatory transcription factor 1 is regulated by diverse stress signals. J Biol Chem 276:25487–25495CrossRefPubMedGoogle Scholar
  40. Song YF, Hogstrand C, Wei CC, Wu K, Pan YX, Luo Z (2017) Endoplasmic reticulum (ER) stress and cAMP/PKA pathway mediated Zn-induced hepatic lipolysis. Environ Pollut 228:256–264CrossRefPubMedGoogle Scholar
  41. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739CrossRefPubMedPubMedCentralGoogle Scholar
  42. Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3:1–12CrossRefGoogle Scholar
  43. Wang F, Kim BE, Petris MJ, Eide DJ (2004) The mammalian Zip5 protein is a zinc transporter that localizes to the basolateral surface of polarized cells. J Biol Chem 279:51433–51441CrossRefPubMedGoogle Scholar
  44. Wei CC, Luo Z, Song YF, Pan YX, Wu K, You WJ (2017) Identification of autophagy related genes LC3 and ATG4 from yellow catfish Pelteobagrus fulvidraco and their transcriptional responses to waterborne and dietborne zinc exposure. Chemosphere 175:228–238CrossRefPubMedGoogle Scholar
  45. Zheng D, Feeney GP, Kille P, Hogstrand C (2008) Regulation of ZIP and ZnT zinc transporters in zebrafish gill: zinc repression of ZIP10 transcription by an intronic MRE cluster. Physiol Genom 34:205–214CrossRefGoogle Scholar
  46. Zheng JL, Luo Z, Liu CX, Chen QL, Tan XY, Zhu QL, Gong Y (2013) Differential effects of acute and chronic zinc (Zn) exposure on hepatic lipid deposition and metabolism in yellow catfish Pelteobagrus fulvidraco. Aquat Toxicol 132–133:173–181CrossRefPubMedGoogle Scholar
  47. Zheng JL, Luo Z, Hu W, Liu CX, Chen QL, Zhu QL, Gong Y (2015) Different effects of dietary Zn deficiency and excess on lipid metabolism in yellow catfish Pelteobagrus fulvidraco. Aquaculture 435:10–17CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture of P.R.C., Fishery CollegeHuazhong Agricultural UniversityWuhanChina

Personalised recommendations