Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
2004-8-6
pubmed:abstractText
Mitochondrial failure to generate ATP can be due to damage to their membranes, which leads to release of solutes, e.g., pyridine nucleotides, from the mitochondrial matrix. We developed a highly sensitive fluorescence assay for detecting a pathologic increase in mitochondrial membrane permeability. The assay is based on coupled enzymatic reactions that produce hydrogen peroxide in the presence of the reduced or oxidized form of nicotinamide adenine dinucleotide (NADH/NAD). The hydrogen peroxide is a substrate for horseradish peroxidase that converts Amplex Red into highly fluorescent Resorufin. The assay is able to detect nanomolar levels of pyridine nucleotides in the medium. Calcium additions to isolated rat brain or liver mitochondria incubated in a potassium-based medium with added enzymes caused osmotic swelling, as detected with light scattering, and production of Resorufin, due to release of NADH/NAD. These events were blocked by cyclosporin A (CsA) or Bongkrekic acid (BKA), inhibitors of the mitochondrial permeability transition (MPT). These results indicate that the NADH/NAD release assay is a simple, reliable, and sensitive method for detecting mitochondrial damage and for screening of compounds that protect mitochondria from injury.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
1385-299X
pubmed:author
pubmed:issnType
Print
pubmed:volume
13
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
176-82
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:15296855-Animals, pubmed-meshheading:15296855-Biological Assay, pubmed-meshheading:15296855-Bongkrekic Acid, pubmed-meshheading:15296855-Brain, pubmed-meshheading:15296855-Cell Membrane Permeability, pubmed-meshheading:15296855-Cyclosporine, pubmed-meshheading:15296855-Drug Evaluation, Preclinical, pubmed-meshheading:15296855-Fluorescent Dyes, pubmed-meshheading:15296855-Histocytochemistry, pubmed-meshheading:15296855-Hydrogen Peroxide, pubmed-meshheading:15296855-Intracellular Membranes, pubmed-meshheading:15296855-Male, pubmed-meshheading:15296855-Mitochondria, pubmed-meshheading:15296855-NAD, pubmed-meshheading:15296855-Neuroprotective Agents, pubmed-meshheading:15296855-Nucleotides, pubmed-meshheading:15296855-Oxazines, pubmed-meshheading:15296855-Pyridines, pubmed-meshheading:15296855-Rats, pubmed-meshheading:15296855-Rats, Wistar, pubmed-meshheading:15296855-Reproducibility of Results
pubmed:year
2004
pubmed:articleTitle
A fluorescence-based technique for screening compounds that protect against damage to brain mitochondria.
pubmed:affiliation
Department of Anesthesiology, School of Medicine, University of Maryland, 685 West Baltimore Street, MSTF 5-34, Baltimore, MD 21201, USA. tkristian@anesthlab.umm.edu
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't