Skip to main content

Advertisement

SpringerLink
Log in

Metallothionein from Pseudosciaena crocea: expression and response to cadmium-induced injury in the testes

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

Metallothioneins (MTs) are a family of stress proteins that are involved in the process of detoxification and anti-oxidation. Previous studies have focused mostly on the expression and functions of MTs in the non-reproductive tissues of aquatic vertebrates. However, there have been only a few reports regarding the functions of MTs in the reproductive tissues of such vertebrates. In order to investigate the function of MTs during spermatogenesis in Pseudosciaena crocea, reverse-transcription polymerase chain reaction (PCR) and rapid amplification of cDNA ends were performed to obtain the P. crocea MT complete cDNA sequence from the total RNA of the testes for the first time. MT was detected in the liver, kidneys, testes, spleen, gill and muscle of P. crocea by tissue-specific expression analysis. Meanwhile, immunohistochemistry staining indicated that the MT protein was localized in germ cells, Sertoli cells and the peripheral connective tissues in P. crocea testes. Furthermore, acute toxicity tests were conducted with cadmium (Cd) to determine the 96 h-medial lethal concentration value. The toxic effects of Cd on the microstructure and ultrastructure of the testes were observed. In addition, the changes in MT mRNA expression levels in the testes after Cd exposure were measured using real-time quantitative PCR. Consequently, we suggest that MTs play an important role in spermatogenesis and testes protection against Cd toxicity in P. crocea.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

Abbreviations

Cd:

Cadmium

Ct:

Threshold cycle

F:

Forward

LC50 :

Medial lethal concentration

MTs:

Metallothioneins

PBS:

Phosphate-buffered saline

R:

Reverse

RACE:

Rapid amplification of cDNA ends

RT-PCR:

Reverse-transcription polymerase chain reaction

SD:

Standard deviation

TD-PCR:

Touchdown PCR

UTRs:

Untranslated regions

References

  • Alessio HM (2000) Lipid peroxidation in healthy and diseases models: influence of different types of exercise. In: Sen CK, Packer L, Hanninen O (eds) Handbook of oxidants and antioxidants in exercise. Elsevier, Amsterdam, pp 115–127

    Google Scholar 

  • ATSDR (1999) Toxicological profile for cadmium (update). Agency for Toxic Substances and Disease Registry, U.S. Department of Health and Human Services, Atlanta

  • Baršytē D, White KN, Lovejoy DA (1999) Cloning and characterization of metallothionein cDNAs in the mussel Mytilus edulis L. digestive gland. Comp Biochem Physiol C: Toxicol Pharmacol 122:287–296

    Article  Google Scholar 

  • Benoff S, Hurley IR, Pergolizzi RG, Goodwin LO (2000) Variation in region IS6 of the L-type voltage-dependent calcium (Ca2+) channel (L-VDCC) α1 subunit in testis and sperm: implications for role of cadmium (Cd2+) in varicocele-associated infertility (VAI). Fertil Steril 74(3):S55–S56

    Article  Google Scholar 

  • Blanco A, Moyano R, Vivo J, Flores-Acuña R, Molina A, Blanco C, Agüera E, Monterde JG (2007) Quantitative changes in the testicular structure in mice exposed to low doses of cadmium. Environ Toxicol Pharmacol 23:96–101

    Article  CAS  Google Scholar 

  • Boujelben M, Ghorbel F, Vincent C, Makni-Ayadi F, Guermazi F, Croute F, El-Feki A (2006) Lipid peroxidation and HSP72/73 expression in rat following cadmium chloride administration: interactions of magnesium supplementation. Exp Toxicol Pathol 57:437–443

    Article  CAS  Google Scholar 

  • Brouwer M, Enghild J, Hoexum-Brouwer T, Thogersen I, Truncai A (1995) Primary structure and tissue-specific expression of blue crab (Callinectes sapidus) metallothionein isoforms. Biochem J 311(2):617–622

    CAS  Google Scholar 

  • Brouwer M, Syring R, Brouwer TH (2002) Role of a copper-specific metallothionein of the blue crab, Callinectes sapidus, in copper metabolism associated with degradation and synthesis of hemocyanin. J Inorg Biochem 88:228–239

    Article  CAS  Google Scholar 

  • Capasso C, Carginale V, Crescenzi O, Maro DD, Parisi E, Spadaccini R, Temussi PA (2003) Solution structure of MT_nc, a novel metallothionein from the Antarctic Fish Notothenia coriiceps. Structure 11:435–443

    Article  CAS  Google Scholar 

  • Chakraborty T, Chatterjee A, Rana A, Srivastawa S, Damodaran S, Chatterjee M (2007) Cell proliferation and hepatocarcinogenesis in rat initiated by diethylnitrosamine and promoted by phenobarbital: potential roles of early DNA damage and liver metallothionein expression. LifeScience 81:489–499

    Article  CAS  Google Scholar 

  • Chavez-Crooker P, Pozo P, Castro H (2003) Cellular localization of calcium, heavy metals, and metallothionein in lobster (Homarus americanus) hepatopancreas. Comp Biochem Physiol C: Toxicol Pharmacol 136(3):213–224

    CAS  Google Scholar 

  • Chen W-Y, John JAC, Lin CH, Lin HF, Wu S-C, Lin C-H, Chang C-Y (2004) Expression of metallothionein gene during embryonic and early larval development in zebrafish. Aquat Toxicol 69:215–227

    Article  CAS  Google Scholar 

  • Chen L, Jin TY, Huang B, Nordberg G, Nordberg M (2006) Critical exposure level of cadmium for elevated urinary metallothionein—an occupational population study in China. Toxicol Appl Pharmacol 215:93–99

    Article  CAS  Google Scholar 

  • Cheuk WK, Chan PC-Y, Chan KM (2008) Cytotoxicities and induction of metallothionein (MT) and metal regulatory element (MRE)-binding transcription factor-1 (MTF-1) messenger RNA levels in the zebrafish (Danio rerio) ZFL and SJD cell lines after exposure to various metal ions. Aquat Toxicol 89:103–112

    Article  CAS  Google Scholar 

  • Cho YS, Lee SY, Kim K-Y, Bang IC, Kim DS, Nam YK (2008) Gene structure and expression of metallothionein during metal exposures in Hemibarbus mylodon. Ecotoxicol Environ Saf 71:125–137

    Article  CAS  Google Scholar 

  • Cho YS, Lee SY, Kim K-Y, Nam YK (2009) Two metallothionein genes from mud loach Misgurnus mizolepis (Teleostei; Cypriniformes): gene structure, genomic organization, and mRNA expression analysis. Comp Biochem Physiol B: Biochem Mol Biol 153:317–326

    Article  Google Scholar 

  • Choi CY, An KW, Nelson ER, Habibi HR (2007) Cadmium affects the expression of metallothionein (MT) and glutathione peroxidase (GPX) mRNA in goldfish, Carassius auratus. Comp Biochem Physiol C: Toxicol Pharmacol 145:595–600

    Google Scholar 

  • Cody CW, Huang PC (1994) Replacement of all α-domain lysines with glutamates reduces metallothionein detoxification function. Biochem Biophys Res Commun 202(2):954–959

    Article  CAS  Google Scholar 

  • Dai SJ, Yin ZQ, Yuan GP, Lu HK, Jia RY, Xu J, Song X, Li L, Shu Y, Liang XX, He CL, Lv C, Zhang W (2013) Quantification of metallothionein on the liver and kidney of rats by subchronic lead and cadmium in combination. Environ Toxicol Pharmacol 36:1207–1216

    Article  CAS  Google Scholar 

  • De SK, Enders GC, Andrews GK (1991) Metallothionein mRNA stability in chicken and mouse cells. BBA 1090(2):223–229

    CAS  Google Scholar 

  • Dohi Y, Shimaoka H, Ikeuchi M, Ohgushi H, Yonemasu K, Minami T (2005) Role of metallothionein isoforms in bone formation processes in rat marrow mesenchymal stem cells in culture. Biol Trace Elem Res 104(1):57–70

    Article  CAS  Google Scholar 

  • Fong L, Tan K, Tran C, Cool J, Scherer MA, Elovaris R, Coyle P, Foster BK, Rofe AM, Xian CJ (2009) Interaction of dietary zinc and intracellular binding protein metallothionein in postnatal bone growth. Bone 44:1151–1162

    Article  CAS  Google Scholar 

  • Gao AB, Wang L, Yuan H (2012) Expression of metallothionein cDNA in a freshwater crab, Sinopotamon yangtsekiense, exposed to cadmium. Exp Toxicol Pathol 64:253–258

    Article  CAS  Google Scholar 

  • Haq F, Mahoney M, Koropatnick J (2003) Signaling events for metallothionein induction. Mutat Res 533:211–226

    Article  CAS  Google Scholar 

  • He L, Girijashanker K, Dalton TP, Reed J, Li H, Soleimani M, Nebert DW (2006) ZIP8, member of the solute-carrier-39 (SLC39) metal transporter family: characterization of transporter properties. Mol Pharmacol 70(1):171–180

    CAS  Google Scholar 

  • Hogstrand C, Haux C (1990) Metallothionein as an indicator of heavy metal exposure in two subtropical fish species. J Exp Mar Biol Ecol 138:69–84

    Article  CAS  Google Scholar 

  • Isani G, Andreani G, Cocchioni F, Fedeli D, Carpené E, Falcioni G (2009) Cadmium accumulation and biochemical responses in Sparus aurata following sub-lethal Cd exposure. Ecotoxicol Environ Saf 72:224–230

    Article  CAS  Google Scholar 

  • Kagi JH, Schaffer A (1988) Biochemistry of metallothionein. Biochemistry 27(23):8509–8515

    Article  CAS  Google Scholar 

  • Kim J-H, Wang S-Y, Kim I-C, Ki J-S, Raisuddin S, Lee J-S, Han K-N (2008) Cloning of a river pufferfish (Takifugu obscurus) metallothionein cDNA and study of its induction profile in cadmium-exposed fish. Chemosphere 71:1251–1259

    Article  CAS  Google Scholar 

  • Kime DE (1999) A strategy for assessing the effects of xenobiotics on fish reproduction. Sci Total Environ 225:3–11

    Article  CAS  Google Scholar 

  • Klaassen CD, Liu J, Diwan BA (2009) Metallothionein protection of cadmium toxicity. Toxicol Appl Pharmacol 238(3):215–220

    Article  CAS  Google Scholar 

  • Kroemer G, Reed JC (2000) Mitochondrial control of cell death. Nat Med 6:513–519

    Article  CAS  Google Scholar 

  • Kusakabe T, Nakajima K, Nakazato K, Suzuki K, Takada H, Satoh T, Oikawa M, Arakawa K, Nagamine T (2008) Changes of heavy metal, metallothionein and heat shock proteins in sertoli cells induced by cadmium exposure. Toxicol In Vitro 22:1469–1475

    Article  CAS  Google Scholar 

  • Langston WJ, Chesman BS, Burt GR, Pope ND, McEvoy J (2002) Metallothionein in liver of eels Anguilla anguilla from the Thames Estuary: an indicator of environmental quality? Mar Environ Res 53:263–293

    Article  CAS  Google Scholar 

  • Le Gall AC, Hydes DJ, Statham PJ, Morley NH, Hunt CL (1999) Processes influencing distributions and concentrations of Cd, Cu, Mn and Ni at the North West European shelf break. Mar Chem 68:97–115

    Article  Google Scholar 

  • Lee JD, Wu SM, Lu LY, Yang YT, Jeng SY (2009) Cadmium concentration and metallothionein expression in prostate cancer and benign prostatic hyperplasia of humans. J Formos Med Assoc 108(7):554–559

    Article  CAS  Google Scholar 

  • Leignel V, Marchand J, Moreau B, Chénais B (2008) Metallothionein genes from hydrothermal crabs (Bythograeidae, Decapoda): characterization, sequence analysis, gene expression and comparison with coastal crabs. Comp Biochem Physiol C: Toxicol Pharmacol 148:6–13

    Google Scholar 

  • Lerebours A, Stentiford GD, Lyons BP, Bignell JP, Derocles SAP, Rotchell JM (2014) Genetic alterations and cancer formation in a European flatfish at sites of different contaminant burdens. Environ Sci Technol 48:10455–10488

    Article  Google Scholar 

  • Li S, Xu JB, Sheng LX (2014) The trans-generation effect during pulsed cadmium exposure: tolerance and induction of hsp70. Ecotoxicol Environ Saf 107:300–305

    Article  CAS  Google Scholar 

  • Lyons-Alcantara M, Mooney R, Lyng F, Cottell D, Mothersill C (1998) The effects of cadmium exposure on the cytology and function of primary cultures from rainbow trout. Cell Biochem Funct 16:1–13

    Article  CAS  Google Scholar 

  • Ma DD, Hou YH, Du LJ, Li N, Xuan RJ, Wang F, Jing WX, Wang L (2013) Oxidative damages and ultrastructural changes in the sperm of freshwater crab Sinopotamon henanense exposed to cadmium. Ecotoxicol Environ Saf 98:244–249

    Article  CAS  Google Scholar 

  • Man AKY, Woo NYS (2008) Upregulation of metallothionein and glucose-6-phosphate dehydrogenase expression in silver sea bream, Sparus sarba exposed to sublethal levels of cadmium. Aquat Toxicol 89:214–221

    Article  CAS  Google Scholar 

  • Mao H, Tan FQ, Wang DH, Zhu JQ, Zhou H, Yang WX (2012a) Expression and function analysis of metallothionein in the testis of stone crab Charybdis japonica exposed to cadmium. Aquat Toxicol 124(125):11–21

    Article  Google Scholar 

  • Mao H, Wang DH, Yang WX (2012b) The involvement of metallothionein in the development of aquatic invertebrate. Aquat Toxicol 110(111):208–213

    Article  Google Scholar 

  • Margoshes M, Vallee BL (1957) A cadmium protein from equine kidney cortex. J Am Chem Soc 79:4813–4814

    Article  CAS  Google Scholar 

  • McKenna IM, Bare RM, Waalkes MP (1996) Metallothionein gene expression in testicular interstitial cells and liver of rats treated with cadmium. Toxicology 107:121–130

    Article  CAS  Google Scholar 

  • Migliarini B, Campisi AM, Maradonna F, Truzzi C, Annibaldi A, Scarponi G, Carnevali O (2005) Effects of cadmium exposure on testis apoptosis in the marine teleost Gobius niger. Gen Comp Endocr 142:241–247

    Article  CAS  Google Scholar 

  • Mignone F, Gissi C, Liuni S, Pesole G (2002) Untranslated regions of mRNAs. Genome Biol 3(3):1–10

    Article  Google Scholar 

  • Moltó E, Bonzón-Kulichenko E, AD Arco, López-Alañón DM, Carrillo O, Gallardo N, Andrés A (2005) Cloning, tissue expression and metal inducibility of an ubiquitous metallothionein from Panulirus argus. Gene 361:140–148

    Article  Google Scholar 

  • Ning Y, Liu XD, Wang ZY, Guo W, Li YY, Xie FJ (2007) A genetic map of large yellow croaker Pseudosciaena crocea. Aquaculture 264:16–26

    Article  CAS  Google Scholar 

  • Oliveira H, Spanò M, Santos C, MDL Pereira (2009) Adverse effects of cadmium exposure on mouse sperm. Reprod Toxicol 28:550–555

    Article  CAS  Google Scholar 

  • Otvos JD, Olafson RW, Armitage IM (1982) Structure of an invertebrate metallothionein from Scylla serrata. J Biol Chem 257:2427–2431

    CAS  Google Scholar 

  • Park JD, Liu YP, Klaassen CD (2001) Protective effect of metallothionein against the toxicity of cadmium and other metals. Toxicology 163:93–100

    Article  CAS  Google Scholar 

  • Ren XY, Zhou Y, Zhang JP, Feng WH, Jiao BH (2003) Metallothionein gene expression under different time in testicular Sertoli and spermatogenic cells of rats treated with cadmium. Reprod Toxicol 17:219–227

    Article  CAS  Google Scholar 

  • Ren F, Jiang H, Sun JL, He L, Li WW, Wang Y, Wang Q (2011) Cloning, characterization, expression, and copper sensitivity of the metallothionein-1 gene in the Chinese mitten crab, Eriocheir sinensis. Mol Biol Rep 38(4):2383–2393

    Article  CAS  Google Scholar 

  • Rigby KE, Stillman MJ (2004) Structural studies of metal-free metallothionein. Biochem Biophys Res Commun 325:1271–1278

    Article  CAS  Google Scholar 

  • Saha P, Mishra D, Chakraborty A, Sudarshan M, Raychaudhuri SS (2008) In vitro radiation induced alterations in heavy metals and metallothionein content in Plantago ovata Forsk. Biol Trace Elem Res 124(3):251–261

    Article  CAS  Google Scholar 

  • Schultze P, Wörgötter E, Braun W, Wagner G, Vašák M, Kägi JHR, Wüthrich K (1988) Conformation of [Cd7]-metallothionein-2 from rat liver in aqueous solution determined by nuclear magnetic resonance spectroscopy. J Mol Biol 203(1):251–268

    Article  CAS  Google Scholar 

  • Scudiero R, Temussi PA, Parisi E (2005) Fish and mammalian metallothioneins: a comparative study. Gene 345:21–26

    Article  CAS  Google Scholar 

  • Shaikh ZA, Vu TT, Zamen K (1999) Oxidative stress as a mechanism of chronic cadmium-induced hepatotoxicity and renal toxicity and protection by antioxidants. Toxicol Appl Pharmacol 154(3):256–263

    Article  CAS  Google Scholar 

  • Siscar R, Koenig S, Torreblanca A, Solé M (2013) The role of metallothionein and selenium in metal detoxification in the liver of deep-sea fish from the NW Mediterranean Sea. Sci Total Environ 466(467):898–905

    Google Scholar 

  • Siu ER, Mruk DD, Porto CS, Cheng CY (2009) Cadmium-induced testicular injury. Toxicol Appl Pharmacol 238:240–249

    Article  CAS  Google Scholar 

  • Soegianto A, Charmantier-Daures M, Trilles JP (1999) Impact of cadmium on the structure of gills and epipodites of the shrimp Penaeus japonicus (Crustacea: Decapoda). Aquat Living Resour 12(1):57–70

    Article  Google Scholar 

  • Son Y-O, Wang X, Hitron JA, Zhang Z, Cheng SP, Budhraja A, Ding SZ, Lee J-C, Shi XL (2011) Cadmium induces autophagy through ROS-dependent activation of the LKB1-AMPK signaling in skin epidermal cells. Toxicol Appl Pharmacol 255:287–296

    Article  CAS  Google Scholar 

  • Susin SA, Lorenzo HK, Zamzami N, Marzo I, Brenner C, Larochette N, Prevost MC, Alzari PM, Kroemer G (1999) Mitochondrial release of caspase-2 and -9 during the apoptotic process. J Exp Med 189:381–394

    Article  CAS  Google Scholar 

  • Tokar EJ, Diwan BA, Waalkes MP (2010) Early life inorganic lead exposure induces testicular teratoma and renal and urinary bladder preneoplasia in adult metallothionein-knockout mice but not in wild type mice. Toxicology 276:5–10

    Article  CAS  Google Scholar 

  • Tremellen K (2008) Oxidative stress and male infertility—a clinical perspective. Hum Reprod Update 14:243–258

    Article  CAS  Google Scholar 

  • Trinchella F, Riggio M, Filosa S, Volpe MG, Parisi E, Scudiero R (2006) Cadmium distribution and metallothionein expression in lizard tissues following acute and chronic cadmium intoxication. Comp Biochem Physiol C: Toxicol Pharmacol 144:272–278

    Article  Google Scholar 

  • Vašák M (2005) Advances in metallothionein structure and functions. J Trace Elem Med Biol 19:13–17

    Article  Google Scholar 

  • Vergani L, Grattarola M, Dondero F, Viarengo A (2003) Expression, purification, and characterization of metallothionein-A from rainbow trout. Protein Expres Purif 27:338–345

    Article  CAS  Google Scholar 

  • Viarengo A, Nott JA (1993) Mechanisms of heavy metal cation homeostasis in marine invertebrates. Comp Biochem Physiol C: Toxicol Pharmacol 104(3):355–372

    Article  Google Scholar 

  • Waisberg M, Joseph P, Hale B, Beyersmann D (2003) Molecular and cellular mechanisms of cadmium carcinogenesis. Toxicology 192:95–117

    Article  CAS  Google Scholar 

  • Wlostowski T, Krasowska A, Laszkiewicz-Tiszczenko B (2000) Dietary cadmium induces histopathological changes despite a sufficient metallothionein level in the liver and kidneys of the bank vole (Clethrionomys glareolus). Comp Biochem Physiol C: Toxicol Pharmacol 126:21–28

    CAS  Google Scholar 

  • Wlostowski T, Krasowska A, Bonda E (2008) Joint effects of dietary cadmium and polychlorinated biphenyls on metallothionein induction, lipid peroxidation and histopathology in the kidneys and liver of blank voles. Ecotoxicol Environ Saf 69:403–410

    Article  CAS  Google Scholar 

  • Xiang DF, Zhu JQ, Jin S, Hu YJ, Tan FQ, Yang WX (2013) Expression and function analysis of metallothionein in the testis of Portunus trituberculatus exposed to cadmium. Aquat Toxicol 140(141):1–10

    Article  Google Scholar 

  • Xu LC, Sun H, Wang SY, Song L, Chang H-C, Wang XR (2005) The roles of metallothionein on cadmium-induced testes damages in Sprague—Dawley rats. Environ Toxicol Pharmacol 20:83–87

    Article  CAS  Google Scholar 

  • Yan CHM, Chan KM (2004) Cloning of zebrafish metallothionein gene and characterization of its gene promoter region in HepG2 cell line. BBA 1679:47–58

    CAS  Google Scholar 

  • Yang QL, Yao CL, Wang ZY (2012) Acute temperature and cadmium stress response characterization of small heat shock protein 27 in large yellow croaker, Larimichthys crocea. Comp Biochem Physiol C: Toxicol Pharmacol 155:190–197

    CAS  Google Scholar 

  • You YL, Lin DJ, Chen LY (2001) Spermatogenesis of teleosts, Pseudosciaena crocea. Zool Res 22(6):461–466 (In Chinese)

    Google Scholar 

Download references

Acknowledgments

We are indebted to Dr. Chun-Yi Hu in Shanghai Institute of Biochemistry and Cell Biology, Dr. Wen-Kai Han in Anhui Normal University, all members of the Sperm Laboratory at Zhejiang University, and all members of our laboratory at Ningbo University for their valuable discussion and inspiring help. This project was financially supported by the National Natural Science Foundation of China (Nos. 31272642 and 41276151), K.C. Wong Magna Fund in Ningbo University, the Scientific and Technical Project of Zhejiang Province (No. 2012C12907-8), the Scientific and Technical Project of Ningbo City (No. 2011C11005), and the Innovation Team Project of Ningbo City (No. 2011B82018).

Conflict of interest

The authors declare that there is no conflict of interest regarding the publication of this paper.

Author information

Authors and Affiliations

  1. School of Marine Sciences, Ningbo University, Ningbo, 315211, Zhejiang, People’s Republic of China

    Zhang Sheng & Jun-Quan Zhu

  2. The Sperm Laboratory, College of Life Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, People’s Republic of China

    Wan-Xi Yang

Authors
  1. Zhang Sheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wan-Xi Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jun-Quan Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jun-Quan Zhu.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sheng, Z., Yang, WX. & Zhu, JQ. Metallothionein from Pseudosciaena crocea: expression and response to cadmium-induced injury in the testes. Ecotoxicology 24, 779–794 (2015). https://doi.org/10.1007/s10646-015-1423-y

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10646-015-1423-y

Keywords

Search

Navigation