Measuring Mitochondrial Substrate Flux in Recombinant Perfringolysin O-Permeabilized Cells
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11120%2F21%3A43921946" target="_blank" >RIV/00216208:11120/21:43921946 - isvavai.cz</a>
Alternative codes found
RIV/00216208:11150/21:10430541
Result on the web
<a href="https://doi.org/10.3791/62902" target="_blank" >https://doi.org/10.3791/62902</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.3791/62902" target="_blank" >10.3791/62902</a>
Alternative languages
Result language
angličtina
Original language name
Measuring Mitochondrial Substrate Flux in Recombinant Perfringolysin O-Permeabilized Cells
Original language description
Mitochondrial substrate flux is a distinguishing characteristic of each cell type, and changes in its components such as transporters, channels, or enzymes are involved in the pathogenesis of several diseases. Mitochondrial substrate flux can be studied using intact cells, permeabilized cells, or isolated mitochondria. Investigating intact cells encounters several problems due to simultaneous oxidation of different substrates. Besides, several cell types contain internal stores of different substrates that complicate results interpretation. Methods such as mitochondrial isolation or using permeabilizing agents are not easily reproducible. Isolating pure mitochondria with intact membranes in sufficient amounts from small samples is problematic. Using non-selective permeabilizers causes various degrees of unavoidable mitochondrial membrane damage. Recombinant perfringolysin O (rPFO) was offered as a more appropriate permeabilizer, thanks to its ability to selectively permeabilize plasma membrane without affecting mitochondrial integrity. When used in combination with microplate respirometry, it allows testing the flux of several mitochondrial substrates with enough replicates within one experiment while using a minimal number of cells. In this work, the protocol describes a method to compare mitochondrial substrate flux of two different cellular phenotypes or genotypes and can be customized to test various mitochondrial substrates or inhibitors.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
30105 - Physiology (including cytology)
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2021
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
JoVE - Journal of Visualized Experiments
ISSN
1940-087X
e-ISSN
—
Volume of the periodical
174
Issue of the periodical within the volume
August
Country of publishing house
US - UNITED STATES
Number of pages
16
Pages from-to
"e62902"
UT code for WoS article
000709869500003
EID of the result in the Scopus database
2-s2.0-85114358485