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Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria
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...
Autores principales: | , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3002942/ https://www.ncbi.nlm.nih.gov/pubmed/21179558 http://dx.doi.org/10.1371/journal.pone.0014359 |
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author | Reiner, Joseph E. Kishore, Rani B. Levin, Barbara C. Albanetti, Thomas Boire, Nicholas Knipe, Ashley Helmerson, Kristian Deckman, Koren Holland |
author_facet | Reiner, Joseph E. Kishore, Rani B. Levin, Barbara C. Albanetti, Thomas Boire, Nicholas Knipe, Ashley Helmerson, Kristian Deckman, Koren Holland |
author_sort | Reiner, Joseph E. |
collection | PubMed |
description | 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. |
format | Text |
id | pubmed-3002942 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-30029422010-12-21 Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria Reiner, Joseph E. Kishore, Rani B. Levin, Barbara C. Albanetti, Thomas Boire, Nicholas Knipe, Ashley Helmerson, Kristian Deckman, Koren Holland PLoS One Research Article 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. Public Library of Science 2010-12-16 /pmc/articles/PMC3002942/ /pubmed/21179558 http://dx.doi.org/10.1371/journal.pone.0014359 Text en This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. https://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. |
spellingShingle | Research Article Reiner, Joseph E. Kishore, Rani B. Levin, Barbara C. Albanetti, Thomas Boire, Nicholas Knipe, Ashley Helmerson, Kristian Deckman, Koren Holland Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria |
title | Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria |
title_full | Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria |
title_fullStr | Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria |
title_full_unstemmed | Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria |
title_short | Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria |
title_sort | detection of heteroplasmic mitochondrial dna in single mitochondria |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3002942/ https://www.ncbi.nlm.nih.gov/pubmed/21179558 http://dx.doi.org/10.1371/journal.pone.0014359 |
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