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Kinetics of the early subcellular distribution of cadmium in rat hepatocytes

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Abstract

The kinetics of the early subcellular distribution of cadmium (Cd) was characterized in primary cultures of rat hepatocytes exposed to 10, 50 and 100 μM Cd in a serum-free WME medium for 10, 30 or 60 min. Our results demonstrate a time- and concentration-dependent increase in Cd content with the highest metal concentration measured in the cytosol, whereas the lowest was observed in the mitochondria. With the exception of early localization in the plasma membrane, Cd concentrations in fractions were characterized by the following decreasing order of magnitude: cytosol > low density molecules nuclei > lysosomes mitochondria. We also found evidence for: (i) a two-step process for Cd distribution in the nuclei and mitochondria; and (ii) a time-dependent ‘slow’ process of transfer from the plasma membrane to the cytosol. Saturation in Cd uptake was observed at 50 μM in most cell fractions at 10 and 30 min, except for the plasma membrane. The lack of apparent saturation for Cd accumulation at 60 min was not related to an increase in metallothionein synthesis. Altogether, our data provide insights into the dynamics of transfer between intracellular compartments, and allow a better identification of the organelles that are the most subjected to Cd toxicity for early exposure conditions.

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References

  • HR Andersen O. Andersen (1988) ArticleTitleEffect of cadmium chloride on hepatic lipid peroxydation in mice Pharmacol Toxicol 63 173–177 Occurrence Handle1:CAS:528:DyaL1cXlvVGitrs%3D Occurrence Handle3186627

    CAS  PubMed  Google Scholar 

  • GK. Andrews (1990) ArticleTitleRegulation of metallothionein gene expression Prog Food Nutr Sci 14 193–258 Occurrence Handle1:STN:280:By6C2MnotVM%3D Occurrence Handle2293243

    CAS  PubMed  Google Scholar 

  • EA Belyaeva VV Glazunov ER Nikitina SM. Korotkov (2001) ArticleTitleBivalent metal ions modulate Cd2+ effects on isolated rat liver mitochondria J Bioenerg Biomembr 33 303–318 Occurrence Handle10.1023/A:1010607307099 Occurrence Handle1:CAS:528:DC%2BD3MXnvF2ntrw%3D Occurrence Handle11710806

    Article  CAS  PubMed  Google Scholar 

  • D Beyersmann S. Hechtenberg (1997) ArticleTitleCadmium, gene regulation, and cellular signaling in mammalian cells Toxicol Appl Pharmacol 144 247–261 Occurrence Handle10.1006/taap.1997.8125 Occurrence Handle1:CAS:528:DyaK2sXjsl2jtLs%3D Occurrence Handle9194408

    Article  CAS  PubMed  Google Scholar 

  • JS Bolduc F Denizeau C. Jumarie (2004) ArticleTitleCadmium-induced mitochondrial membrane potential dissipation does not necessarily require cytololic oxidative stress: studies using rhodamine-123 fluorescence unquenching Toxicol Sci 77 299–306 Occurrence Handle10.1093/toxsci/kfh015 Occurrence Handle1:CAS:528:DC%2BD2cXht1KltbY%3D Occurrence Handle14600273

    Article  CAS  PubMed  Google Scholar 

  • MM. Bradford (1976) ArticleTitleA rapid and sensitive method for the quantitation of microgram quantitites of protein utilizing the principle of protein-dye binding Anal Biochem 72 248–254 Occurrence Handle1:CAS:528:DyaE28XksVehtrY%3D Occurrence Handle942051

    CAS  PubMed  Google Scholar 

  • P Chavez-Crooker N Garrido GA. Ahearn (2003) ArticleTitleCopper transport by lobster (Homarus americanus) hepatopancreatic lysosomes Comp Biochem Physiol 135 IssueIDC 107–118

    Google Scholar 

  • G. Cherian (1980) ArticleTitleThe synthesis of metallothionein and cellular adaptation to metal toxicity in primary rat kidney epithelial cell cultures Toxicology 17 225–231 Occurrence Handle10.1016/0300-483X(80)90098-0 Occurrence Handle1:CAS:528:DyaL3MXkt1aksw%3D%3D Occurrence Handle7210007

    Article  CAS  PubMed  Google Scholar 

  • IN. Chou (1989) ArticleTitleDistinct cytosqueletal injuries induced by arsenic, cadmium, cobalt, chromic, and nickel compounds Biomed Environ Sci 2 358–365 Occurrence Handle1:STN:280:By%2BC3cvmsVY%3D Occurrence Handle2604903

    CAS  PubMed  Google Scholar 

  • CC Conrad CA Walter A Richardson MA Hanes DT. Grabowski (1997) ArticleTitleCadmium toxicity and distribution in metallothionein-I and metallothionein-II deficient transgenic mice J Toxicol Environ Health 52 527–543 Occurrence Handle10.1080/009841097159502 Occurrence Handle1:CAS:528:DyaK2sXns1ejtbg%3D Occurrence Handle9397184

    Article  CAS  PubMed  Google Scholar 

  • KG Danielson S Ohi PC. Huang (1982) ArticleTitleImmunochemical detection of metallothionein in specific epithelial cells of rat organs Proc Natl Acad Sci USA 79 2301–2304 Occurrence Handle1:CAS:528:DyaL38Xhslyms7o%3D Occurrence Handle7048317

    CAS  PubMed  Google Scholar 

  • SR Davis RJ. Cousins (2000) ArticleTitleMetallothionein expression in animals: a physiological perspective on function J Nutr 130 1085–1088

    Google Scholar 

  • RE Dudley DJ Swoboda CD. Klaassen (1984) ArticleTitleTime course of cadmium-induced ultrastructural changes in rat liver Toxicol Appl Pharmacol 76 50–160 Occurrence Handle10.1016/0041-008X(84)90038-3

    Article  Google Scholar 

  • SA El-Maraghy MZ Gad AT Fahim MA. Hamdy (2001) ArticleTitleEffect of cadmium and aluminium intake on the antioxydant status and lipid peroxidation in rat tissues J Biochem Mol Toxicol 15 207–214 Occurrence Handle10.1002/jbt.18 Occurrence Handle1:CAS:528:DC%2BD3MXlslSgtL8%3D Occurrence Handle11673849

    Article  CAS  PubMed  Google Scholar 

  • PL Goering MP Waalkes CD Klaassen (1995) Toxicology of cadmium RA Goyer MG Cherian (Eds) Toxicology of Metals, Biochemical Aspects Springer Berlin 189–214

    Google Scholar 

  • PL Goering CD. Klaassen (1983) ArticleTitleAltered subcellular distribution of cadmium following cadmium pretreatment: possible mechanism of tolerance to cadmium-induced lethality Toxicol Appl Pharmacol 70 195–203 Occurrence Handle10.1016/0041-008X(83)90095-9 Occurrence Handle1:CAS:528:DyaL3sXlslSrtLg%3D Occurrence Handle6623465

    Article  CAS  PubMed  Google Scholar 

  • Z Guan B Kukoyi P Feng MC Kennedy RB Franklin LC. Costello (2003) ArticleTitleKinetic identification of a mitochondrial zinc uptake transport process in prostate cells J Inorg Biochem 97 199–206 Occurrence Handle10.1016/S0162-0134(03)00291-5 Occurrence Handle1:CAS:528:DC%2BD3sXnsVKltbc%3D Occurrence Handle14512198

    Article  CAS  PubMed  Google Scholar 

  • T Hamada A Tanimoto S Iwai H Fujiwara Y. Sasaguri (1994) ArticleTitleCytopathological changes induced by cadmium-exposure in canine proximal tubular cells: a cytochemical and ultrastructural study Nephron 68 104–111 Occurrence Handle1:CAS:528:DyaK2cXmt1eru7g%3D Occurrence Handle7991019

    CAS  PubMed  Google Scholar 

  • D Horky I Lauschova J Illek A Pechova M. Sindelar (2002) ArticleTitleDistribution of exogenous heavy metals in the hepatocytes of calves: a morphometric study Microsc Res Tech 56 451–453 Occurrence Handle10.1002/jemt.10044 Occurrence Handle1:CAS:528:DC%2BD38Xis1amtbg%3D Occurrence Handle11921346

    Article  CAS  PubMed  Google Scholar 

  • CJ Hsiao SR. Stapleton (2004) ArticleTitleCharacterization of Cd-induced molecular events prior to cellular damage in primary rat hepatocytes in culture: activation of the stress activated signal protein JNK and transcription factor AP-1 J Biochem Mol Toxicol 18 133–142 Occurrence Handle10.1002/jbt.20018 Occurrence Handle1:CAS:528:DC%2BD2cXlslelsL8%3D Occurrence Handle15252869

    Article  CAS  PubMed  Google Scholar 

  • SH Jeong SS Habeebu CD Klaassen (2000) ArticleTitleCadmium decreases gap junctional intercellular communication in mouse liver Toxicol Sci 57 156–166 Occurrence Handle10.1093/toxsci/57.1.156 Occurrence Handle1:CAS:528:DC%2BD3cXmslylsbw%3D Occurrence Handle10966522

    Article  CAS  PubMed  Google Scholar 

  • T Koizumi H Shirakura H Kumagai H Tatsumoto KT Suzuki (1996) ArticleTitleMechanisms of cadmium-induced cytotoxicity in rat hepatocytes: cadmium-induced active oxygen-related permeability changes of the plasma membrane Toxicology 114 125–134 Occurrence Handle10.1016/S0300-483X(96)03477-4 Occurrence Handle1:CAS:528:DyaK28XmvFelurk%3D Occurrence Handle8947611

    Article  CAS  PubMed  Google Scholar 

  • S Koizumi H Yamada (2003) ArticleTitleDNA microarray analysis of altered gene expression in cadmium-exposed human cells J Occup Health 45 331–334 Occurrence Handle10.1539/joh.45.331 Occurrence Handle1:CAS:528:DC%2BD2cXjt1WqsQ%3D%3D Occurrence Handle14676411

    Article  CAS  PubMed  Google Scholar 

  • A Lemarié D Lagadic-Gossmann C Morzadec N Allain O Fardel L Vernhet (2004) ArticleTitleCadmium induces caspase-independent apoptosis in liver Hep3B cells: role for calcium in signalling oxidative stress-related impairment of mitochondria and relocation of endonuclease G and apoptosis-inducing factor Free Radic Biol Med 36 1517–1531 Occurrence Handle10.1016/j.freeradbiomed.2004.03.020 Occurrence Handle15182854

    Article  PubMed  Google Scholar 

  • JJ Lemasters (1999) ArticleTitleMechanisms of hepatic toxicity: part V. Necrapoptosis and the mitochondrial permeability transition: shared pathways to necrosis and apoptosis Am J Physiol Gastrointest Liver Physiol 276 G1–G6 Occurrence Handle1:CAS:528:DyaK1MXntlGitg%3D%3D

    CAS  Google Scholar 

  • L Li J Kaplan (2004) ArticleTitleA mitochondrial-vacuolar signaling pathway in yeast that affects iron and copper metabolism J Biol Chem 279 33651–33661

    Google Scholar 

  • W Li HM Kagan IN. Chou (1994) ArticleTitleAlterations in cytoskeletal organization and homeostasis of cellular thiols in cadmium-resistant cells Toxicol Appl Pharmacol 126 114–123 Occurrence Handle10.1006/taap.1994.1097 Occurrence Handle1:CAS:528:DyaK2cXjtFOmsLg%3D Occurrence Handle8184421

    Article  CAS  PubMed  Google Scholar 

  • J Liu WC Kershanw CD. Klaassen (1991) ArticleTitleThe protective effect of metallothionein on the toxicity of various metals in rat primary hepatocyte culture Toxicol Appl Pharmacol 107 27–34 Occurrence Handle10.1016/0041-008X(91)90327-B Occurrence Handle1:CAS:528:DyaK3MXps1Cisw%3D%3D Occurrence Handle1987657

    Article  CAS  PubMed  Google Scholar 

  • Y Liu J Liu CD. Klaassen (2001) ArticleTitleMetallothionein-null and wild type mice show similar cadmium absorption and tissue distribution following oral cadmium administration Toxicol Appl Pharmacol 175 253–259 Occurrence Handle10.1006/taap.2001.9244 Occurrence Handle1:CAS:528:DC%2BD3MXmsl2nu7k%3D Occurrence Handle11559024

    Article  CAS  PubMed  Google Scholar 

  • D Manca M Lefevre B Trottier et al. (1992) ArticleTitleMicro method for determination of cadmium in tissues and slurried samples by use of flameless atomic absorption spectrometry Microchem J 46 249–258 Occurrence Handle10.1016/0026-265X(92)90044-4 Occurrence Handle1:CAS:528:DyaK38Xmt1Ortbk%3D

    Article  CAS  Google Scholar 

  • M Margoshes BL. Vallee (1957) ArticleTitleA cadmium protein from equine kidney cortex J Am Chem Soc 79 4813–4814 Occurrence Handle10.1021/ja01574a064 Occurrence Handle1:CAS:528:DyaG2sXpvFKhsQ%3D%3D

    Article  CAS  Google Scholar 

  • RLW Messer LC. Lucas (2002) ArticleTitleLocalization of metallic ions within gingival fibroblast subcellular fractions J Biomed Mater Res 59 466–472 Occurrence Handle10.1002/jbm.1262 Occurrence Handle1:CAS:528:DC%2BD38Xjt1Srsg%3D%3D Occurrence Handle11774304

    Article  CAS  PubMed  Google Scholar 

  • RR Misra JE Page GT Smith MP Waalkes A. Dipple (1998) ArticleTitleEffect of cadmium exposure on background and anti-5 methylchrysene-1,2-dihydrodiol 3,4-epoxide-induced mutagenesis in the supF gene of pS189 in human Ad293 cells Chem Res Toxicol 11 211–216 Occurrence Handle10.1021/tx970183b Occurrence Handle1:CAS:528:DyaK1cXhslarsLs%3D Occurrence Handle9544619

    Article  CAS  PubMed  Google Scholar 

  • CA Mizzen NJ Cartel WH Yu PE Fraser DR. McLaclan (1996) ArticleTitleSensitive detection of metallothionein-1, −2 and −3 in tissue homogenates by immunoblotting: a method for enhanced membrane transfer and retention J Biochem Biophys Methods 32 77–83 Occurrence Handle10.1016/0165-022X(95)00044-R Occurrence Handle1:CAS:528:DyaK28XjtlSrtbo%3D Occurrence Handle8796480

    Article  CAS  PubMed  Google Scholar 

  • L. Muller (1986) ArticleTitleConsequences of cadmium toxicity in rat hepatocytes: mitochondrial dysfunction and lipid peroxydation Toxicology 40 285–295 Occurrence Handle10.1016/0300-483X(86)90061-2 Occurrence Handle1:STN:280:BimA3MnnslQ%3D Occurrence Handle3750329

    Article  CAS  PubMed  Google Scholar 

  • S Nakamura T Kawata A Nakayama K Kubo T Minami H. Sakurai (2004) ArticleTitleImplication of the differential roles of metallothionein 1 and 2 isoforms in the liver of rats as determined by polyacrylamide-coated capillary zone electrophoresis Biochem Biophys Res Comm 320 1193–1198 Occurrence Handle10.1016/j.bbrc.2004.06.085 Occurrence Handle1:CAS:528:DC%2BD2cXls1eis7k%3D Occurrence Handle15249216

    Article  CAS  PubMed  Google Scholar 

  • GF Nordberg T Jin M. Nordberg (1994) ArticleTitleSubcellular targets of cadmium nephrotoxicity: cadmium binding to renal membrane proteins in animals with or without protective metallothionein synthesis Environ Health Perspect 102 191–194 Occurrence Handle1:CAS:528:DyaK2cXmvFCmtb0%3D

    CAS  Google Scholar 

  • RD. Palmiter (1998) ArticleTitleThe elusive function of metallothioneins Proc Natl AcadSci USA 95 8428–8430 Occurrence Handle10.1073/pnas.95.15.8428 Occurrence Handle1:CAS:528:DyaK1cXkvFamtbo%3D

    Article  CAS  Google Scholar 

  • MJ Perez AI. Cederbaum (2003) ArticleTitleMetallothionein 2A induction by zinc protects HEPG2 cells against CYP2E1-dependent toxicity Free Radic Biol Med 34 443–455 Occurrence Handle10.1016/S0891-5849(02)01302-3 Occurrence Handle1:CAS:528:DC%2BD3sXptFGiuw%3D%3D Occurrence Handle12566070

    Article  CAS  PubMed  Google Scholar 

  • P Pourahmad PJ O’Brien F Jokar B. Daraei (2003) ArticleTitleCarcinogenic metal induced sites of reactive oxygen species formation in hepatocytes Toxicol in Vitro 17 803–810 Occurrence Handle10.1016/S0887-2333(03)00123-1 Occurrence Handle1:CAS:528:DC%2BD3sXos12lt78%3D Occurrence Handle14599481

    Article  CAS  PubMed  Google Scholar 

  • WC Prozialeck GB Grunwald M Dey KR Reuhl AR. Parrish (2002) ArticleTitleCadherins and NCAM as potential targets in metal toxicity Toxicol Appl Pharmacol 182 255–265 Occurrence Handle10.1006/taap.2002.9422 Occurrence Handle1:CAS:528:DC%2BD38XmtFSisL0%3D Occurrence Handle12183105

    Article  CAS  PubMed  Google Scholar 

  • W Rau F Planas-Bohne DM. Taylor (1987) ArticleTitleInfluence of several chelating agents on the distribution and binding of cadmium in rats Hum Toxicol 6 451–458 Occurrence Handle1:CAS:528:DyaL1cXjtlagsw%3D%3D Occurrence Handle3692490

    CAS  PubMed  Google Scholar 

  • XY Ren Y Zhou JP Zhang WH Feng BH. Jiaco (2003) ArticleTitleMetallothionein gene expression under different time in testicular Sertoli and spermatogenic cells of rats treated with cadmium Reproductive Toxicol 17 219–227 Occurrence Handle10.1016/S0890-6238(02)00151-X Occurrence Handle1:CAS:528:DC%2BD3sXitV2nsbY%3D

    Article  CAS  Google Scholar 

  • PO. Seglen (1976) ArticleTitlePreparation of isolated rat liver cells Methods Cell Biol 13 29–83 Occurrence Handle1:CAS:528:DyaE28XkslCgtbY%3D Occurrence Handle177845

    CAS  PubMed  Google Scholar 

  • ZA Shaikh TT Vu K. Zaman (1999) ArticleTitleOxidative stress as a mechanism of chronic cadmium-induced hepatotoxicity and renal toxicity and protection by antioxidants Toxicol Appl Pharmacol 145 256–263 Occurrence Handle10.1006/taap.1998.8586

    Article  Google Scholar 

  • N Shiraishi JF Hochadel TP Coogan J Koropatnick MP. Waalkes (1995) ArticleTitleSensitivity to cadmium-induced genotoxocoty in rat testicular cells is associated with minimal expression of the metallothionein gene Toxicol Appl Pharmacol 130 229–236 Occurrence Handle10.1006/taap.1995.1028 Occurrence Handle1:CAS:528:DyaK2MXjvFCntbY%3D Occurrence Handle7871536

    Article  CAS  PubMed  Google Scholar 

  • IA Simpson DR Yver PJ Hissin et al. (1983) ArticleTitleInsulin-stimulated translocation of glucose transporters in the isolated rat adipose cells: characterization of subcellular fractions Biochim Biophys Acta 763 393–407 Occurrence Handle10.1016/0167-4889(83)90101-5 Occurrence Handle1:CAS:528:DyaL2cXmt1amsQ%3D%3D Occurrence Handle6360220

    Article  CAS  PubMed  Google Scholar 

  • KN Tzirogiannis GI Panoutsopoulos MD Demonakou et al. (2003) ArticleTitleTime-course of cadmium-induced acute hepatotoxicity in the rat liver: the role of apoptosis Arch Toxicol 77 694–701 Occurrence Handle10.1007/s00204-003-0499-y Occurrence Handle1:CAS:528:DC%2BD3sXptleks7k%3D Occurrence Handle13680093

    Article  CAS  PubMed  Google Scholar 

  • MH Vasconcelos SC Tam JE Hesketh M Reid JH. Beattie (2002) ArticleTitleMetal- and tissue-dependent relationship between metallothionein mRNA and protein Toxicol Appl Pharmacol 182 91–97 Occurrence Handle10.1006/taap.2002.9428 Occurrence Handle1:CAS:528:DC%2BD38XlsFWqtr8%3D Occurrence Handle12140172

    Article  CAS  PubMed  Google Scholar 

  • T Von Zglinicki C Edwall E Ostlund B Lind M Norberg NR Ringertz J. Wroblewski (1992) ArticleTitleVery low cadmium concentrations stimulate DNA synthesis and cell growth J Cell Sci 103 1073–1081 Occurrence Handle1:CAS:528:DyaK3sXhs1alt7s%3D Occurrence Handle1487490

    CAS  PubMed  Google Scholar 

  • M Waisberg P Joseph B Hale D. Beyersmann (2003) ArticleTitleMolecular and cellular mechanisms of cadmium carcinogenesis Toxicology 192 95–117 Occurrence Handle10.1016/S0300-483X(03)00305-6 Occurrence Handle1:CAS:528:DC%2BD3sXot1Kjt7w%3D Occurrence Handle14580780

    Article  CAS  PubMed  Google Scholar 

  • K. Waku (1984) ArticleTitleThe chemical form of cadmium in subcellular fractions following cadmium exposure Environ Health Perspect 54 37–44 Occurrence Handle1:CAS:528:DyaL2cXksV2ktrY%3D Occurrence Handle6376094

    CAS  PubMed  Google Scholar 

  • W Wätjen D. Beyersmann (2004) ArticleTitleCadmium-induced apoptosis in C6 glioma cells: influence of oxidative stress Biometals 17 65–78 Occurrence Handle10.1023/A:1024405119018 Occurrence Handle14977363

    Article  PubMed  Google Scholar 

  • Wells WW, Seyfred MA, Smith CD, Sakai M. (1987). Measurement of subcellular sites of polyphosphoinositide metabolism in isolated rat hepatocytes. In Conn PM, Means AR, eds. Methods in enzymology: Cellular Regulators Vol. 141, part B, 92–99

  • T Wolstowski A. Krasowska (1999) ArticleTitleSubcellular distribution of metallothionein and cadmium in the liver and kidney of bank voles (Clethrionomys glareolus) exposed to dietary cadmium Biometals 12 173–179 Occurrence Handle10.1023/A:1009285820760 Occurrence Handle10406086

    Article  PubMed  Google Scholar 

  • T Zhou G Zhou W Song N Eguchi W Lu E Lundin T Jin G. Nordberg (1999) ArticleTitleCadmium-induced apoptosis and changes in expression of p53, c-jun and MT-1 genes in testes and ventral prostate of rats Toxicology 142 1–13 Occurrence Handle10.1016/S0300-483X(99)00115-8 Occurrence Handle1:CAS:528:DyaK1MXns1Smsbw%3D Occurrence Handle10647914

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Catherine Jumarie.

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Deceased March 25, 2004.

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Diep, P.T.N., Denizeau, F. & Jumarie, C. Kinetics of the early subcellular distribution of cadmium in rat hepatocytes. Biometals 18, 255–267 (2005). https://doi.org/10.1007/s10534-005-1538-3

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