• Rev Med Chir Soc Med Nat Iasi · Jan 2012

    High-resolution respirometry with multiple substrates titration in permeabilized myocardial fibers.

    • Oana Duicu, Dorina Gheorgheosu, Nicoleta Mirica, Simona Trancotă, Dehelean Cristina, O Firă-Mladinescu, and Danina Muntean.
    • Department of Pathophysiology, "Victor Babeş" University of Medicine and Pharmacy Timisoara, Romania.
    • Rev Med Chir Soc Med Nat Iasi. 2012 Jan 1; 116 (1): 207-13.

    AbstractThe present study was purported to standardize the high-resolution respirometry technique for the analysis of oxidative phosphorylation (OXPHOS) in saponin-skinned cardiac fibers. Preparation of permeabilized myocardial fibers allows the assessment of respiratory function of the entire population of mitochondria in their normal intracellular position. Adult male rat ventricular bundles were permeabilized with saponin (50 microg/ml) and samples (1-3 mg wet weight) were transferred into the Oxygraph-2k (OROBOROS Instr., Austria) chambers containing air saturated incubation buffer in order to measure Complex I (CI) and II (CII) dependent respiration. The following values (expressed in pmol O2 x s(-1) x mg(-1)) were obtained: CI_LEAK, 67.18 +/- 5.12 (CI dependent basal respiration, after glutamate and malate addition); CI_P, 247.37 +/- 49.90 (CI_OXPHOS state after ADP addition); CI_Pc, 252.036 +/- 53.13 (the intactness of the outer mitochondrial membrane assessed after cytochrome c addition); CI+II_P, 342.90 +/- 62.48 (maximum OXPHOS state obtained after succinate addition by activating convergent electron flow from CI+II into the Q junction of the electron transport system (ETS); CII_P, 302.26 +/- 50.16 (CII dependent OXPHOS state obtained after the addition of a CI inhibitor rotenone, with the subsequent entry of electrons from CII only into the Q junction); ETS capacity, 331.11 +/- 62.39 (uncoupled respiration). The standardized technique will allow the systematic characterization of mitochondrial respiratory dysfunction associated with several cardiac pathologies in both animals and humans.

      Pubmed     Copy Citation     Plaintext  

      Add institutional full text...

    Notes

     
    Knowledge, pearl, summary or comment to share?
    300 characters remaining
    help        
    You can also include formatting, links, images and footnotes in your notes
    • Simple formatting can be added to notes, such as *italics*, _underline_ or **bold**.
    • Superscript can be denoted by <sup>text</sup> and subscript <sub>text</sub>.
    • Numbered or bulleted lists can be created using either numbered lines 1. 2. 3., hyphens - or asterisks *.
    • Links can be included with: [my link to pubmed](http://pubmed.com)
    • Images can be included with: ![alt text](https://bestmedicaljournal.com/study_graph.jpg "Image Title Text")
    • For footnotes use [^1](This is a footnote.) inline.
    • Or use an inline reference [^1] to refer to a longer footnote elseweher in the document [^1]: This is a long footnote..

    hide…