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Evolution of metallotionein isoforms complexes in hepatic cells of Mus musculus along cadmium exposure

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Abstract

Characterization of Cd-binding proteins has great analytical interest due to the high toxicity of Cd to living organisms. Metallothioneins (MTs), as Cd(II)-binding proteins are of increasing interest, since they form very stable Cd chelates and are involved in many detoxification processes. In this work, inductively coupled plasma octopole reaction cell mass spectrometry and nanospray ionization time-of-flight mass spectrometry were used in parallel and combined with two-dimensional chromatography: size exclusion followed by reversed-phase high performance liquid chromatography, to study metal complexes of MT isoforms produced in hepatic cytosols of Mus musculus during exposure experiments to Cd. Exposure experiments were carried out by subcutaneous injection of a growing dose of the toxic element ranging from 0.1 to 1.0 mg of Cd per kg of body weight per day during 10 days. A control group and three exposure groups at days 2, 6 and 10 of exposure were studied, and different cadmium, copper and zinc complexes with MTs isoforms were isolated and characterized from the two most exposed groups. The results allow gaining insight into the mechanisms involved in metal detoxification by MTs, showing the changes in the stoichiometry of metal complexes–MTs along cadmium exposure.

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References

  • Adam V, Fabrik I, Eckschlager T, Stiborova M, Trnkova L, Kizek R (2010) Vertebrate metallothioneins as target molecules for analytical techniques. Trends Anal Chem 29:409–418

    Article  CAS  Google Scholar 

  • Andón B, Barbosa J, Sanz-Nebot V (2006) Separation and characterization of rabbit liver apothioneins by capillary electrophoresis coupled to electrospray ionization time-of-flight mass spectrometry. Electrophoresis 27:3661–3670

    Article  PubMed  Google Scholar 

  • Brzoska MM, Moniuszko-Jakoniuk J, Pilat-Marcinkiewicz B, Sawicki B (2003) Liver and kidney function and histology in rats exposed to cadmium and ethanol. Alcohol 38:2–10

    CAS  Google Scholar 

  • Cai L, Li XK, Song Y, Cherian MG (2005) Essentiality, toxicology and chelation therapy of zinc and copper. Curr Med Chem 12:2753–2763

    Article  PubMed  CAS  Google Scholar 

  • Chassaige H, Lobinski R (1998) Speciation of metal complexes with biomolecules by reversed- phase HPLC wit ion-spray and inductively. Fresenius J Anal Chem 361:267–273

    Article  Google Scholar 

  • Chassaigne H, Lobinski R (1998) Characterization of horse kidney metallothionein isoforms by electrospray MS and reversed-phase HPLC-electrospray MS. Analyst 123:2125–2130

    Article  PubMed  CAS  Google Scholar 

  • Chassaigne H, Lobinski R (1999) Detection of artefacts and peak identification in reversed-phase HPLC of metallothioneins by electrospray mass spectrometry. Talanta 48:109–118

    Article  PubMed  CAS  Google Scholar 

  • Chassaigne H, Łobinski R (1998) Characterization of metallothionein isoforms by reversed-phase high-performance liquid chromatography with on-line post-column acidification and electrospray mass spectrometric detection. J Chromatogr A 829:127–136

    Article  PubMed  CAS  Google Scholar 

  • el Jihen H, Imed M, Fatima H, Abdelhamid K (2009) Protective effects of selenium (Se) and zinc (Zn) on cadmium (Cd) toxicity in the liver of the rat: effects on the oxidative stress. Ecotoxicol Environ Saf 72:1559–1564

    Article  CAS  Google Scholar 

  • Fargosava A (1998) Comparative acute toxicity of Cu2+, Cu+, Mn2+, Mo6+, Ni2+, and V5+ to Chironomus plumosus larvae and Tubifex tubifex worms. Biologia 53:315–319

    Google Scholar 

  • Gehrig PM, You C, Dallinger R, Gruber C, Brouwer M, Kagi JHR, Hunziker PE (2000) Electrospray ionization mass spectrometry of zinc, cadmium, and copper metallothioneins: evidence for metal-binding cooperativity. Protein Sci 9:395–402

    Article  PubMed  CAS  Google Scholar 

  • Gonzalez-Fernandez M, García-Barrera T, Arias-Borrego A, Jurado J, Pueyo C, López-Barea J, Gómez-Ariza JL (2009) Metallomics integrated with proteomics in deciphering metal-related environmental issues. Biochimie 91:1311–1317

    Article  PubMed  CAS  Google Scholar 

  • Gonzalez-Fernandez M, García-Sevillano MA, Jara-Biedma R, García-Barrera T, Vioque A, López-Barea J, Pueyo C, Gómez-Ariza JL (2011) Size characterization of metal species in liver and brain from free-living (Mus spretus) and laboratory (Mus musculus) mice by SEC-ICP-MS: application to environmental contamination assessment. J Anal At Spectrom 26:141–149

    Article  CAS  Google Scholar 

  • Hathout Y, Reynolds KJ, Szilagyi Z, Fenselau C (2002) Metallothionein dimers studied by nano-spray mass spectrometry. J Inorg Biochem 88:119–122

    Article  PubMed  CAS  Google Scholar 

  • Jarup L, Akesson A (2009) Current status of cadmium as an environmental health problem. Toxicol Appl Pharmacol 238:201–208

    Article  PubMed  Google Scholar 

  • Joohyun L, Donghwan S, Won-Yong S, Inhwan H, Youngsook L (2004) Arabidopsis metallothioneins 2a and 3 enhance resistance to cadmium when expressed in Vicia faba guard cells. Plant Mol Biol 54:805–815

    Article  Google Scholar 

  • Jurczuk M, Brzoska MM, Moniuszko-Jakoniuk J (2004) Antioxidant enzymes activity and lipid peroxidation in liver and kidney of rats exposed to cadmium and ethanol. Food Chem Toxicol 42:429–438

    Article  PubMed  CAS  Google Scholar 

  • Jurczuk M, Moniuszko-Jakoniuk J, Brzoska MM, Roszczenko A (2005) Vitamins E and C concentrations in the liver and kidney of rats exposed to cadmium and ethanol. Pol J Environ Stud 14:599–604

    CAS  Google Scholar 

  • Klassen CD, Liu J, Choudhuri S (1999) Metallothionein: an intracellular protein to protect against cadmium toxicity. Ann Rev Pharmacol Toxicol 39:267–294

    Article  Google Scholar 

  • Koyu A, Gokcimen A, Ozguner F (2006) Evaluation of the effects of cadmium on rat liver. Mol Cell Biochem 284:81–85

    Article  PubMed  CAS  Google Scholar 

  • Martelli A, Rousselet E, Dycke C, Moulis JM (2006) Cadmium toxicity in animal cells by interference with essential metals. Biochimie 88:1807–1814

    Article  PubMed  CAS  Google Scholar 

  • Merrifield ME, Huang Z, Kille P, Stillman MJ (2002) Copper speciation in the alpha and beta domains of recombinant human metallothionein by electrospray ionization mass spectrometry. J Inorg Biochem 88:153–172

    Article  PubMed  CAS  Google Scholar 

  • Moniuszko-Jakoniuk J, Jurczuk M, Brzoska MM, Rogalska J, Galazyn-Sidorczuk M (2005) Involvement of some low-molecular thiols in the destructive mechanism of cadmium and ethanol action on rat livers and kidneys. Pol Environ Stud 14:483–489

    CAS  Google Scholar 

  • Mounicou S, Polec K, Chassaigne H, Potin-Gautier M, Lobinski R (2000) Characterization of metal complexes with metallothioneins by capillary zone electrophoresis (CZE) with ICP-MS and electrospray (ES)-MS detection. J Anal At Spectrom 15:635–642

    Article  CAS  Google Scholar 

  • Mounicou S, Szpunar J, Lobinski R (2009) Metallomics: the concept and methodology. Chem Soc Rev 38:1119–1138

    Article  PubMed  CAS  Google Scholar 

  • Mounicou S, Ouerdane L, L’Azou B, Passagne I, Ohayon-Courtès C, Szpunar J, Lobinski R (2010) Identification of metallothionein subisoforms in HPLC using accurate mass and online sequencing by electrospray hybrid linear ion trap-orbital ion trap mass spectrometry. Anal Chem 82:6947–6957

    Article  PubMed  CAS  Google Scholar 

  • Murugavel P, Pari L (2007) Effects of diallyl tetrasulfide on cadmium-induced oxidative damage in the liver of rats. Hum Exp Toxicol 26:527–534

    Article  PubMed  CAS  Google Scholar 

  • Nawrot TS, Staessen JA, Roels HA (2010) Cadmium exposure in the population: from health risks to strategies of prevention. Biometals 23:769–782

    Article  PubMed  CAS  Google Scholar 

  • Nischwitz V, Michalke B, Kettrup A (2003) Identification and quantification of metallothionein isoforms and superoxide dismutase in spiked liver extracts using HPLC-ESI-MS offline coupling and HPLC-ICP-MS online coupling. Anal Bioanal Chem 375:145–156

    PubMed  CAS  Google Scholar 

  • Nordberg GF (2009) Historical perspectives on cadmium toxicology. Toxicol Appl Pharmacol 238:192–200

    Article  PubMed  CAS  Google Scholar 

  • Podhorska-Okołów M, Dziegiel P, Dolińska-Krajewska B, Dumańska M, Cegielski M, Jethon Z, Rossini K, Carraro U, Zabel M (2006) Expression of metallothionein in renal tubules of rats exposed to acute and endurance exercise. Folia Histochem Cytobiol 44(3):195–200

    PubMed  Google Scholar 

  • Poléc K, Perez-Calvo M, Garcia-Arribas O, Szpunar J, Ribas-Ozonas B, Lobinski R (2002) Investigation of metal complexes with metallothionein in rat tissues by hyphenated techniques. J Inorg Biochem 88:197–206

    Article  PubMed  Google Scholar 

  • Prange A, Schaumlöffel D (2002) Hyphenated techniques for the characterization and quantification of metallothionein isoforms: review-analytical and bioanalytical. Chemistry 373:441–453

    CAS  Google Scholar 

  • Rana SVS (2008) Metals and apoptosis: recent developments. J Trace Elem Med Biol 22:262–284

    Article  PubMed  CAS  Google Scholar 

  • Renugadevi J, Milton-Prabu S (2010) Cadmium-induced hepatotoxicity in rats and the protective effect of naringenin. Exp Toxicol Pathol 62:171–181

    Article  PubMed  CAS  Google Scholar 

  • Richards MP, Steele NC (1987) Isolation and quantitation of metallothionein isoforms using reversed-phase high-performance liquid chromatography. J Chromatogr 402:243–256

    Article  PubMed  CAS  Google Scholar 

  • Rogival D, Van Campenhout K, Goenaga Infante H, Hearn R, Scheirs J, Blust R (2007) Induction and metal speciation of metallothionein in wood mice (Apodemus sylvaticus) along a metal pollution gradient. Environ Toxicol Chem 26:506–514

    Article  PubMed  CAS  Google Scholar 

  • Ryvolova M, Krizkova S, Adam V, Beklova M, Trnkova L, Hubalek J, Kizek R (2011) Analytical methods for metallothionein detection. Curr Anal Chem 7:243–261

    CAS  Google Scholar 

  • Shen J, Huang Z, Zhuang Z, Wang X, Lee FCS (2005) Investigation of mercury metallothionein complexes in tissues of rat after oral intake of HgCl2. Appl Organometal Chem 19:140–146

    Article  CAS  Google Scholar 

  • Shimoda R, Achanzar WE, Qu W, Nagamine T, Takagi H, Mori M, Waalkes MP (2003) Metallothionein is a potential negative regular of apoptosis. Toxicol Sci 73:294–300

    Article  PubMed  CAS  Google Scholar 

  • Souza V, Del Escobar CM, Bucio L, Hernandez E, Gutierrez-Ruiz MC (2004) Zinc pretreatment prevents hepatic stellate cells from cadmium-produced oxidative damage. Cell Biol Toxicol 20:241–251

    Article  PubMed  CAS  Google Scholar 

  • Srivastava RC, Ahmad I, Kaur G, Hasan SK (1988) Alterations in the metabolism of endogenous trace metals due to cadmium, manganese and nickel-effect of partial hepatectomy. J Environ Sci Health A 23:95–101

    Article  Google Scholar 

  • Stillman MJ, Thomas D, Trevithick C, Guo X, Siu M (2000) Circular dichroism, kinetic and mass spectrometric studies of copper(I) and mercury(II) binding to metallothionein. J Inorg Biochem 79:11–19

    Article  PubMed  CAS  Google Scholar 

  • Strouhal M, Kizek R, Vacek J, Trnková L, Nemec M (2003) Electrochemical study of heavy metals and metallothionein in yeast Yarrowia lipolytica. Bioelectrochemistry 60:29–36

    Article  PubMed  CAS  Google Scholar 

  • Suzuki KT, Sunaga H, Aoki Y, Yamamura M (1983) Gel permeation, ion-exchange and reversed-phase columns for separation of metallothioneins by high-performance liquid chromatography-atomic absorption spectrophotometry. J Chromatogr 281:159–166

    Article  PubMed  CAS  Google Scholar 

  • Szpunar J (2005) Advances in analytical methodology for bioinorganic speciation analysis: metallomics, metalloproteomics and heteroatom-tagged proteomics and metabolomics. Analyst 130:442–465

    Article  PubMed  CAS  Google Scholar 

  • Templeton DM, Liu Y (2010) Multiple roles of cadmium in cell death and survival. Chem Biol Interact 188:267–275

    Article  PubMed  CAS  Google Scholar 

  • Tzirogiannis KN, Panoutsopoulos GI, Demonakou MD, Hereti RI, Alexandropoulou KN, Basayannis AC, Mykoniatis MG (2003) Time-course of cadmium induced acute hepatotoxicity in the rat liver: the role of apoptosis. Arch Toxicol 77:694–701

    Article  PubMed  CAS  Google Scholar 

  • Van der Oost R, Beyer J, Vermeulen NPE (2003) Fish bioaccumulation and biomarkers in environmental risk assessment: a review. Environ Toxicol Pharmacol 13:57–149

    Article  PubMed  Google Scholar 

  • Webb D, Gagnon MM, Rose T (2005) Metabolic enzyme activities in black bream (Acanthopagrus butcheri) from the Swan-Canning Estuary, Western Australia. Comp Biochem Phys C 141:356–365

    Article  Google Scholar 

  • Yang MS, Lai KP, Cheng KY, Wong CKC (2000) Changes in endogenous Zn and Cu distribution in different cytosolic protein fractions in mouse liver after administration of a single sublethal dose of CdCl2. Toxicology 154:103–111

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The authors thank the Projects CTM2012-38720-C03-01 (Ministerio de Economia y Competitividad-Spain), P08-FQM-03554 and P09-FQM-04659 (Consejería de Innovación, Andalusian government). R. González-Domínguez thanks to Ministerio de Ciencia e Innovación for a predoctoral grant. R. Jara-Biedma thanks to Consejería de Innovación. Junta de Andalucia for a predoctoral grant.

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Correspondence to T. García-Barrera or J. L. Gómez-Ariza.

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Jara-Biedma, R., González-Dominguez, R., García-Barrera, T. et al. Evolution of metallotionein isoforms complexes in hepatic cells of Mus musculus along cadmium exposure. Biometals 26, 639–650 (2013). https://doi.org/10.1007/s10534-013-9636-0

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