Measurement of H2O2 within living Drosophila during aging using a ratiometric mass spectrometry probe targeted to the mitochondrial matrix.

TitleMeasurement of H2O2 within living Drosophila during aging using a ratiometric mass spectrometry probe targeted to the mitochondrial matrix.
Publication TypeJournal Article
Year of Publication2011
AuthorsCochemé, HM, Quin, C, McQuaker, SJ, Cabreiro, F, Logan, A, Prime, TA, Abakumova, I, Patel, JV, Fearnley, IM, James, AM, Porteous, CM, Smith, RAJ, Saeed, S, Carré, JE, Singer, M, Gems, D, Hartley, RC, Partridge, L, Murphy, MP
JournalCell Metab
Volume13
Issue3
Pagination340-50
Date Published2011 Mar 2
ISSN1932-7420
KeywordsAging, Animals, Chromatography, High Pressure Liquid, Drosophila, Hydrogen Peroxide, Mitochondria, Organophosphorus Compounds, Phenols, Tandem Mass Spectrometry
Abstract

Hydrogen peroxide (H(2)O(2)) is central to mitochondrial oxidative damage and redox signaling, but its roles are poorly understood due to the difficulty of measuring mitochondrial H(2)O(2) in vivo. Here we report a ratiometric mass spectrometry probe approach to assess mitochondrial matrix H(2)O(2) levels in vivo. The probe, MitoB, comprises a triphenylphosphonium (TPP) cation driving its accumulation within mitochondria, conjugated to an arylboronic acid that reacts with H(2)O(2) to form a phenol, MitoP. Quantifying the MitoP/MitoB ratio by liquid chromatography-tandem mass spectrometry enabled measurement of a weighted average of mitochondrial H(2)O(2) that predominantly reports on thoracic muscle mitochondria within living flies. There was an increase in mitochondrial H(2)O(2) with age in flies, which was not coordinately altered by interventions that modulated life span. Our findings provide approaches to investigate mitochondrial ROS in vivo and suggest that while an increase in overall mitochondrial H(2)O(2) correlates with aging, it may not be causative.

DOI10.1016/j.cmet.2011.02.003
Alternate JournalCell Metab.
Citation Key10.1016/j.cmet.2011.02.003
PubMed ID21356523
PubMed Central IDPMC4413513
Grant ListBB/D020786/1 / / Biotechnology and Biological Sciences Research Council / United Kingdom
MC_U105663142 / / Medical Research Council / United Kingdom
/ / Wellcome Trust / United Kingdom
/ / Medical Research Council / United Kingdom
/ / Biotechnology and Biological Sciences Research Council / United Kingdom
/ / Department of Health / United Kingdom