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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

  • Czech description

Classification

  • Type

    J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database

  • CEP classification

  • 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