Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria

Published

December 17, 2010

Author(s)

Joseph E. Reiner, Rani Kishore, Barbara C. Levin, Thomas Albanetti, Nicholas Boire, Ashley Knipe, Kristian Helmerson, Koren Deckman

Abstract

Background Mitochondrial DNA (mtDNA) genome mutations can lead to energy and respiratory-related disorders like myoclonic epilepsy with ragged red fiber disease (MERRF), mitochondrial myopathy, encephalopathy, lactic acidosis and stroke (MELAS) syndrome, and Leber s hereditary optic neuropathy (LHON). It is not well understood what effect the distribution of mutated mtDNA throughout the mitochondrial matrix has on the development of mitochondrial-based disorders. Insight into this complex sub-cellular heterogeneity may further our understanding of the development of mitochondria-related diseases. Methodology This work describes a method for isolating individual mitochondria from single cells and performing molecular analysis on that single mitochondrion s DNA. An optical tweezer extracts a single mitochondrion from a lysed human HL-60 cell. Then a micron-sized femtopipette tip captures the mitochondrion for subsequent analysis. Multiple rounds of conventional DNA amplification and standard sequencing methods enable the detection of a heteroplasmic mixture in the mtDNA from a single mitochondrion. Significance Molecular analysis of mtDNA from the individually extracted mitochondrion demonstrates that a heteroplasmy is present in single mitochondria at various ratios consistent with the 50/50 heteroplasmy ratio found in single cells that contain multiple mitochondria.

Citation

PLoS One

Volume

5

Issue

12

Pub Type

Journals

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Keywords

nanobiotechnology, optical tweezers, mitochondria, DNA technologies

Citation

Reiner, J. , Kishore, R. , Levin, B. , Albanetti, T. , Boire, N. , Knipe, A. , Helmerson, K. and Deckman, K. (2010), Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria, PLoS One, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=904019 (Accessed December 3, 2024)