skip to content
Measurement of mitochondrial respiration in permeabilized murine neuroblastoma (N-2alpha) cells, a simple and rapid in situ assay to investigate mitochondrial toxins.
ClosePreview this item
Checking...

Measurement of mitochondrial respiration in permeabilized murine neuroblastoma (N-2alpha) cells, a simple and rapid in situ assay to investigate mitochondrial toxins.

Author: SJ Steyn Affiliation: Department of Biochemistry, North West University, Potchefstroom, South Africa 2520.; DJ Pieterse; LJ Mienie; Van der Schyf CJ
Edition/Format: Article Article : English
Publication:Journal of biochemical and biophysical methods, 2005 Jan 31; 62(1): 25-40
Summary:
Most mitochondria-based methods used to investigate toxins require the use of relatively large amounts of material and hence compromised sensitivity in assay. We adopted procedures from methods initially developed to diagnose mitochondrial encephalomyopathies and unified these into a single assay. Eukaryotic cell membranes are selectively permeabilized with digitonin to render a system in which mitochondrial  Read more...
Rating:

(not yet rated) 0 with reviews - Be the first.

&AllPage.SpinnerRetrieving;

Find a copy in the library

&AllPage.SpinnerRetrieving; Finding libraries that hold this item...

Details

Document Type: Article
All Authors / Contributors: SJ Steyn Affiliation: Department of Biochemistry, North West University, Potchefstroom, South Africa 2520.; DJ Pieterse; LJ Mienie; Van der Schyf CJ
ISSN:0165-022X
Language Note: English
Unique Identifier: 110344382
Awards:

Abstract:

Most mitochondria-based methods used to investigate toxins require the use of relatively large amounts of material and hence compromised sensitivity in assay. We adopted procedures from methods initially developed to diagnose mitochondrial encephalomyopathies and unified these into a single assay. Eukaryotic cell membranes are selectively permeabilized with digitonin to render a system in which mitochondrial respiration can be measured rapidly and with considerable sensitivity. Mitochondria remain intact, uninjured, and in their natural environment where mitochondrial respiration can be measured in situ under physiologically relevant conditions. This approach furthermore allows measurement of toxin effects on individual mitochondrial complexes. Numerous compounds at varying concentrations can be screened for mitochondrial toxicity, while the site of mitochondrial inhibition can be determined simultaneously. We used this assay to investigate, in murine neuroblastoma (N-2alpha) cells, the mitochondrial inhibitory properties of the parkinsonian-inducing proneurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and its neurotoxic monoamine oxidase-B (MAO-B)-generated metabolite, the 1-methyl-4-phenylpyridinium species (MPP(+)). Within the time frame of each measurement (15 min), MPTP (< or = 1 mM) did not interfere with in situ mitochondrial respiration. As expected, MPP(+) was found to be a potent Complex I inhibitor but surprisingly also found to inhibit Complex IV. Optimized conditions for performing this assay are provided.

Reviews

User-contributed reviews
Retrieving GoodReads reviews...
Retrieving DOGObooks reviews...

Tags

Be the first.
Confirm this request

You may have already requested this item. Please select Ok if you would like to proceed with this request anyway.

Linked Data


Primary Entity

<http://www.worldcat.org/oclc/110344382> # Measurement of mitochondrial respiration in permeabilized murine neuroblastoma (N-2alpha) cells, a simple and rapid in situ assay to investigate mitochondrial toxins.
    a schema:Article, schema:CreativeWork ;
    library:oclcnum "110344382" ;
    rdfs:comment "949 $l Research Support, Non-U.S. Gov't" ;
    rdfs:comment "949 $l Journal Article" ;
    schema:contributor <http://experiment.worldcat.org/entity/work/data/4352767666#Person/pieterse_dj> ; # Pieterse DJ
    schema:contributor <http://experiment.worldcat.org/entity/work/data/4352767666#Person/mienie_lj> ; # Mienie LJ
    schema:contributor <http://experiment.worldcat.org/entity/work/data/4352767666#Person/van_der_schyf_cj> ; # Van der Schyf CJ
    schema:creator <http://experiment.worldcat.org/entity/work/data/4352767666#Person/steyn_sj> ; # Steyn SJ
    schema:description "Most mitochondria-based methods used to investigate toxins require the use of relatively large amounts of material and hence compromised sensitivity in assay. We adopted procedures from methods initially developed to diagnose mitochondrial encephalomyopathies and unified these into a single assay. Eukaryotic cell membranes are selectively permeabilized with digitonin to render a system in which mitochondrial respiration can be measured rapidly and with considerable sensitivity. Mitochondria remain intact, uninjured, and in their natural environment where mitochondrial respiration can be measured in situ under physiologically relevant conditions. This approach furthermore allows measurement of toxin effects on individual mitochondrial complexes. Numerous compounds at varying concentrations can be screened for mitochondrial toxicity, while the site of mitochondrial inhibition can be determined simultaneously. We used this assay to investigate, in murine neuroblastoma (N-2alpha) cells, the mitochondrial inhibitory properties of the parkinsonian-inducing proneurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and its neurotoxic monoamine oxidase-B (MAO-B)-generated metabolite, the 1-methyl-4-phenylpyridinium species (MPP(+)). Within the time frame of each measurement (15 min), MPTP (< or = 1 mM) did not interfere with in situ mitochondrial respiration. As expected, MPP(+) was found to be a potent Complex I inhibitor but surprisingly also found to inhibit Complex IV. Optimized conditions for performing this assay are provided." ;
    schema:exampleOfWork <http://worldcat.org/entity/work/id/4352767666> ;
    schema:isPartOf <http://worldcat.org/issn/0165-022X#62_1> ;
    schema:name "Measurement of mitochondrial respiration in permeabilized murine neuroblastoma (N-2alpha) cells, a simple and rapid in situ assay to investigate mitochondrial toxins." ;
    schema:pageStart "25" ;
    schema:productID "110344382" ;
    wdrs:describedby <http://www.worldcat.org/title/-/oclc/110344382> ;
    .


Related Entities

<http://experiment.worldcat.org/entity/work/data/4352767666#Person/van_der_schyf_cj> # Van der Schyf CJ
    a schema:Person ;
    schema:name "Van der Schyf CJ" ;
    .

<http://worldcat.org/issn/0165-022X>
    a schema:Periodical ;
    rdfs:label "Journal of biochemical and biophysical methods" ;
    schema:issn "0165-022X" ;
    .

<http://www.worldcat.org/title/-/oclc/110344382>
    a genont:InformationResource, genont:ContentTypeGenericResource ;
    schema:about <http://www.worldcat.org/oclc/110344382> ; # Measurement of mitochondrial respiration in permeabilized murine neuroblastoma (N-2alpha) cells, a simple and rapid in situ assay to investigate mitochondrial toxins.
    schema:dateModified "2017-08-27" ;
    void:inDataset <http://purl.oclc.org/dataset/medline> ;
    .


Content-negotiable representations

Close Window

Please sign in to WorldCat 

Don't have an account? You can easily create a free account.