Neurotoxicity Research

, Volume 34, Issue 1, pp 74–78 | Cite as

Association Study Between Metallothionein-3 Protein Polymorphisms and Autism

  • MingXia Yu
  • Tao Cao
  • Dan Yu
  • Fusheng Huang


Genetic susceptibility to high mercury body burden has been suggested as an autism risk factor in children. Metallothionein III (MT3) is the brain-specific form of the metallothionein family, which plays a key role in metal metabolism. We therefore looked for genetic variations in the MT3 gene that might increase the predisposition to autism. DNA was extracted from 132 autistic children and 132 age and gender-matched unrelated controls. All the samples were analyzed for nine single nucleotide polymorphisms (SNPs) with minor allele frequency > 10% in the MT3 gene. The mRNA levels of MT3 in white blood cells were evaluated by real-time PCR. We did not detect any association between these MT3 polymorphisms and the mRNA levels of MT3. We did not detect any association between MT3 polymorphisms and autism risk. However, we detected four novel MT3 SNPs that are not in the human SNP database. The clinical importance of these SNPs needs further investigation. Our data suggest that MT3 gene polymorphisms are not associated with autism.


Mercury Autism Metallothionein III (MT3) SNP Association study 



The authors greatly thank Prof. Lin Jun for her assistance of diagnosis and are grateful to the subjects and their families for participation and collaboration.


This study was supported by the National Natural Science Foundation of China (Grant No: 81,171,669).

Compliance with Ethical Standards

This study was approved by the local Ethics Committees and Hospital Ethics Committee. Written informed consent was obtained from all participating individuals or parents and/or legal guardians.

Conflicts of Interest

The authors declare that they have no conflict of interest.


  1. Aschner M (1996) The functional significance of brain metallothioneins. FASEB J 10(10):1129–1136CrossRefPubMedGoogle Scholar
  2. Aschner M, Ceccatelli S (2010) Are neuropathological conditions relevant to ethylmercury exposure? Neurotox Res 18(1):59–68. CrossRefPubMedGoogle Scholar
  3. Aschner M, Lorscheider FL, Cowan KS, Conklin DR, Vimy MJ, Lash LH (1997) Metallothionein induction in fetal rat brain and neonatal primary astrocyte cultures by in utero exposure to elemental mercury vapor (Hg0). Brain Res 778(1):222–232. CrossRefPubMedGoogle Scholar
  4. Aschner M, Syversen T, Souza DO, Rocha JB (2006) Metallothioneins: mercury species-specific induction and their potential role in attenuating neurotoxicity. Exp Biol Med (Maywood) 231(9):1468–1473. CrossRefGoogle Scholar
  5. Barrett JC, Fry B, Maller J, Daly MJ (2005) Haploview: analysis and visualization of LD and haplotype maps. Bioinformatics 21(2):263–265. CrossRefPubMedGoogle Scholar
  6. Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55(4):611–622. CrossRefPubMedGoogle Scholar
  7. Chaste P, Leboyer M (2012) Autism risk factors: genes, environment, and gene-environment interactions. Dialogues Clin Neurosci 14(3):281–292PubMedPubMedCentralGoogle Scholar
  8. Garrecht M, Austin DW (2011) The plausibility of a role for mercury in the etiology of autism: a cellular perspective. Toxicol Environ Chem 93(5–6):1251–1273. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Hidalgo J, Aschner M, Zatta P, Vasak M (2001) Roles of the metallothionein family of proteins in the central nervous system. Brain Res Bull 55(2):133–145. CrossRefPubMedGoogle Scholar
  10. Koressaar T, Remm M (2007) Enhancements and modifications of primer design program Primer3. Bioinformatics 23(10):1289–1291. CrossRefPubMedGoogle Scholar
  11. Laura V, Cristina L, Paola R, Luisa AM, Shyti G, Edvige V et al (2011) Metals, metallothioneins and oxidative stress in blood of autistic children. Res Autism Spect Dis 5:286–293CrossRefGoogle Scholar
  12. Manso Y, Carrasco J, Comes G, Meloni G, Adlard PA, Bush AI, Vasak M, Hidalgo J (2012) Characterization of the role of metallothionein-3 in an animal model of Alzheimer’s disease. Cell Mol Life Sci 69(21):3683–3700. CrossRefPubMedGoogle Scholar
  13. Owens SE, Summar ML, Ryckman KK, Haines JL, Reiss S, Summar SR, Aschner M (2011) Lack of association between autism and four heavy metal regulatory genes. Neurotoxicology 32(6):769–775. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Park JD, Liu Y, Klaassen CD (2001) Protective effect of metallothionein against the toxicity of cadmium and other metals(1). Toxicology 163(2–3):93–100. CrossRefPubMedGoogle Scholar
  15. Russo AF (2008) Anti-metallothionein IgG and levels of metallothionein in autistic families. Swiss Med Wkly 138(5–6):70–77PubMedGoogle Scholar
  16. Russo AJ (2009) Anti-metallothionein IgG and levels of metallothionein in autistic children with GI disease. Drug Healthc Patient Saf 1:1–8CrossRefPubMedPubMedCentralGoogle Scholar
  17. Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 3(6):1101–1108. CrossRefPubMedGoogle Scholar
  18. Schultz ST (2010) Does thimerosal or other mercury exposure increase the risk for autism? A review of current literature. Acta Neurobiol Exp (Wars) 70(2):187–195Google Scholar
  19. Singh VK, Hanson J (2006) Assessment of metallothionein and antibodies to metallothionein in normal and autistic children having exposure to vaccine-derived thimerosal. Pediatr Allergy Immunol 17(4):291–296. CrossRefPubMedGoogle Scholar
  20. Sun X, Ding H, Hung K, Guo B (2000) A new MALDI-TOF based mini-sequencing assay for genotyping of SNPS. Nucleic Acids Res 28(12):E68. CrossRefPubMedPubMedCentralGoogle Scholar
  21. Veenstra-Vanderweele J, Christian SL, Cook EH Jr (2004) Autism as a paradigmatic complex genetic disorder. Annu Rev Genomics Hum Genet 5(1):379–405. CrossRefPubMedGoogle Scholar
  22. Walker SJ, Segal J, Aschner M (2006) Cultured lymphocytes from autistic children and non-autistic siblings up-regulate heat shock protein RNA in response to thimerosal challenge. Neurotoxicology 27(5):685–692. CrossRefPubMedGoogle Scholar
  23. Wang Y, Goodrich JM, Gillespie B, Werner R, Basu N, Franzblau A (2012) An investigation of modifying effects of metallothionein single-nucleotide polymorphisms on the association between mercury exposure and biomarker levels. Environ Health Perspect 120(4):530–534. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Woods JS, Heyer NJ, Russo JE, Martin MD, Pillai PB, Farin FM (2013) Modification of neurobehavioral effects of mercury by genetic polymorphisms of metallothionein in children. Neurotoxicol Teratol 39:36–44. CrossRefPubMedGoogle Scholar
  25. Yamada M, Hayashi S, Hozumi I, Inuzuka T, Tsuji S, Takahashi H (1996) Subcellular localization of growth inhibitory factor in rat brain: light and electron microscopic immunohistochemical studies. Brain Res 735(2):257–264. CrossRefPubMedGoogle Scholar
  26. Yoshida M, Watanabe C, Satoh M, Yasutake A, Sawada M, Ohtsuka Y, Akama Y, Tohyama C (2004) Susceptibility of metallothionein-null mice to the behavioral alterations caused by exposure to mercury vapor at human-relevant concentration. Toxicol Sci 80(1):69–73. CrossRefPubMedGoogle Scholar
  27. Yoshida M, Watanabe C, Kishimoto M, Yasutake A, Satoh M, Sawada M, Akama Y (2006) Behavioral changes in metallothionein-null mice after the cessation of long-term, low-level exposure to mercury vapor. Toxicol Lett 161(3):210–218. CrossRefPubMedGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Center for Gene DiagnosisZhongnan Hospital of Wuhan UniversityWuhanPeople’s Republic of China
  2. 2.Department of LaboratoryHubei Cancer HospitalWuhanPeople’s Republic of China

Personalised recommendations