Analysis of Human Mitochondrial DNA Content by Southern Blotting and Nonradioactive Probe Hybridization. Issue 1 (14th April 2019)

• Record Type: Journal Article
• Authors: Wheeler, Joel H.; Young, Carolyn K. J.; Young, Matthew J.
• Editors: Bus, James S.; Costa, Lucio G.; Hodgson, Ernest; Lawrence, David A.; Reed, Donald J.
• Is Part Of: Current protocols in toxicology. Volume 80:Issue 1(2019)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2019-04-14
• Subjects: digoxigenin‐labeling of DNA probes -- Drug‐induced mitochondrial toxicity -- immunodetection -- mitochondrial DNA (mtDNA) depletion -- Southern blotting; Toxicology -- Laboratory manuals; Toxicology -- methods; Toxicology; Laboratory Manual; Electronic books; Laboratory manuals; 571.95028
• DOI: 10.1002/cptx.75
• Languages: English
• ISSNs: 1934-9254

Abstract

Abstract: A single cell can contain several thousand copies of the mitochondrial DNA genome or mtDNA. Tools for assessing mtDNA content are necessary for clinical and toxicological research, as mtDNA depletion is linked to genetic disease and drug toxicity. For instance, mtDNA depletion syndromes are typically fatal childhood disorders that are characterized by severe declines in mtDNA content in affected tissues. Mitochondrial toxicity and mtDNA depletion have also been reported in human immunodeficiency virus–infected patients treated with certain nucleoside reverse transcriptase inhibitors. Further, cell culture studies have demonstrated that exposure to oxidative stress stimulates mtDNA degradation. Here we outline a Southern blot and nonradioactive digoxigenin‐labeled probe hybridization method to estimate mtDNA content in human genomic DNA samples. © 2019 by John Wiley & Sons, Inc.