Abstract
There is increasing evidence that exposure to air pollutants is associated with human disease and may act through epigenetic modification of the nuclear genome, but there have been few publications describing their impact upon the mitochondrial genome. Mitochondrial DNA may be more susceptible to pollutant-induced changes via increased oxidative stress in the cell, and therefore this field of research is of growing interest. Many techniques employed to study DNA methylation of the nuclear genome are also applicable to mitochondrial epigenetic studies. In this chapter, we describe a protocol for the isolation of mitochondrial DNA from peripheral blood samples and the analysis of 5-methylcytosine content by bisulfite pyrosequencing.
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References
Zhang X, Lin S, Funk WE, Hou L (2013) Environmental and occupational exposure to chemicals and telomere length in human studies. Occup Environ Med 70:743–749
Dosunmu R, Alashwal H, Zawia NH (2012) Genome-wide expression and methylation profiling in the aged rodent brain due to early-life Pb exposure and its relevance to aging. Mech Ageing Dev 133:435–443
Bodin J, Bolling AK, Becher R, Kuper F, Lovik M, Nygaard UC (2013) Transmaternal bisphenol a exposure accelerates diabetes type 1 development in NOD mice. Toxicol Sci 137(2):311–323
Pan WC, Seow WJ, Kile ML, Hoffman EB, Quamruzzaman Q, Rahman M et al (2013) Association of low to moderate levels of arsenic exposure with risk of type 2 diabetes in Bangladesh. Am J Epidemiol 178:1563–1570
Loane C, Pilinis C, Lekkas TD, Politis M (2013) Ambient particulate matter and its potential neurological consequences. Rev Neurosci 24:323–335
Silins I, Hogberg J (2011) Combined toxic exposures and human health: biomarkers of exposure and effect. Int J Environ Res Public Health 8:629–647
Byun HM, Motta V, Panni T, Bertazzi PA, Apostoli P, Hou L et al (2013) Evolutionary age of repetitive element subfamilies and sensitivity of DNA methylation to airborne pollutants. Part Fibre Toxicol 10:28
Tarantini L, Bonzini M, Apostoli P, Pegoraro V, Bollati V, Marinelli B et al (2009) Effects of particulate matter on genomic DNA methylation content and iNOS promoter methylation. Environ Health Perspect 117:217–222
Bollati V, Baccarelli A, Hou L, Bonzini M, Fustinoni S, Cavallo D et al (2007) Changes in DNA methylation patterns in subjects exposed to low-dose benzene. Cancer Res 67:876–880
Byun HM, Baccarelli AA (2014) Environmental exposure and mitochondrial epigenetics: study design and analytical challenges. Hum Genet 133(3):247–257
Davidson SM, Duchen MR (2007) Endothelial mitochondria: contributing to vascular function and disease. Circ Res 100:1128–1141
Iacobazzi V, Castegna A, Infantino V, Andria G (2013) Mitochondrial DNA methylation as a next-generation biomarker and diagnostic tool. Mol Genet Metab 110:25–34
Chestnut BA, Chang Q, Price A, Lesuisse C, Wong M, Martin LJ (2011) Epigenetic regulation of motor neuron cell death through DNA methylation. J Neurosci 31:16619–16636
Manev H, Dzitoyeva S, Chen H (2012) Mitochondrial DNA: a blind spot in neuroepigenetics. Biomol Concepts 3:107–115
Dzitoyeva S, Chen H, Manev H (2012) Effect of aging on 5-hydroxymethylcytosine in brain mitochondria. Neurobiol Aging 33:2881–2891
Sun C, Reimers LL, Burk RD (2011) Methylation of HPV16 genome CpG sites is associated with cervix precancer and cancer. Gynecol Oncol 121:59–63
Byun HM, Panni T, Motta V, Hou L, Nordio F, Apostoli P et al (2013) Effects of airborne pollutants on mitochondrial DNA methylation. Part Fibre Toxicol 10:18
Shock LS, Thakkar PV, Peterson EJ, Moran RG, Taylor SM (2011) DNA methyltransferase 1, cytosine methylation, and cytosine hydroxymethylation in mammalian mitochondria. Proc Natl Acad Sci U S A 108:3630–3635
Infantino V, Castegna A, Iacobazzi F, Spera I, Scala I, Andria G et al (2011) Impairment of methyl cycle affects mitochondrial methyl availability and glutathione level in Down’s syndrome. Mol Genet Metab 102:378–382
Song L, James SR, Kazim L, Karpf AR (2005) Specific method for the determination of genomic DNA methylation by liquid chromatography-electrospray ionization tandem mass spectrometry. Anal Chem 77:504–510
Hong EE, Okitsu CY, Smith AD, Hsieh CL (2013) Regionally specific and genome-wide analyses conclusively demonstrate the absence of CpG methylation in human mitochondrial DNA. Mol Cell Biol 33:2683–2690
Mikeska T, Bock C, El-Maarri O, Hubner A, Ehrentraut D, Schramm J et al (2007) Optimization of quantitative MGMT promoter methylation analysis using pyrosequencing and combined bisulfite restriction analysis. J Mol Diagn 9:368–381
Aparicio A, North B, Barske L, Wang X, Bollati V, Weisenberger D et al (2009) LINE-1 methylation in plasma DNA as a biomarker of activity of DNA methylation inhibitors in patients with solid tumors. Epigenetics 4:176–184
Hazkani-Covo E, Zeller RM, Martin W (2010) Molecular poltergeists: mitochondrial DNA copies (numts) in sequenced nuclear genomes. PLoS Genet 6:e1000834
Li LC, Dahiya R (2002) MethPrimer: designing primers for methylation PCRs. Bioinformatics 18:1427–1431
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Byun, HM., Barrow, T.M. (2015). Analysis of Pollutant-Induced Changes in Mitochondrial DNA Methylation. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine. Methods in Molecular Biology, vol 1265. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2288-8_19
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DOI: https://doi.org/10.1007/978-1-4939-2288-8_19
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2287-1
Online ISBN: 978-1-4939-2288-8
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