High resolution respirometry to assess function of mitochondria in native homogenates of human heart muscle
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11110%2F20%3A10403734" target="_blank" >RIV/00216208:11110/20:10403734 - isvavai.cz</a>
Alternative codes found
RIV/00216208:11120/20:43919628 RIV/00064173:_____/20:N0000167 RIV/00023001:_____/20:00079153 RIV/00064165:_____/20:10403734
Result on the web
<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=UbPz52jMwM" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=UbPz52jMwM</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1371/journal.pone.0226142" target="_blank" >10.1371/journal.pone.0226142</a>
Alternative languages
Result language
angličtina
Original language name
High resolution respirometry to assess function of mitochondria in native homogenates of human heart muscle
Original language description
Impaired myocardial bioenergetics is a hallmark of many cardiac diseases. There is a need of a simple and reproducible method of assessment of mitochondrial function from small human myocardial tissue samples. In this study we adopted high-resolution respirometry to homogenates of fresh human cardiac muscle and compare it with isolated mitochondria. We used atria resected during cardiac surgery (n = 18) and atria and left ventricles from brain-dead organ donors (n = 12). The protocol we developed consisting of two-step homogenization and exposure of 2.5% homogenate in a respirometer to sequential addition of 2.5 mM malate, 15 mM glutamate, 2.5 mM ADP, 10 μM cytochrome c, 10 mM succinate, 2.5 μM oligomycin, 1.5 μM FCCP, 3.5 μM rotenone, 4 μM antimycin and 1 mM KCN or 100 mM Sodium Azide. We found a linear dependency of oxygen consumption on oxygen concentration. This technique requires < 20 mg of myocardium and the preparation of the sample takes <20 min. Mitochondria in the homogenate, as compared to subsarcolemmal and interfibrillar isolated mitochondria, have comparable or better preserved integrity of outer mitochondrial membrane (increase of respiration after addition of cytochrome c is up to 11.7+-1.8% vs. 15.7+-3.1%, p<0.05 and 11.7+-3.5%, p = 0.99, resp.) and better efficiency of oxidative phosphorylation (Respiratory Control Ratio = 3.65+-0.5 vs. 3.04+-0.27, p<0.01 and 2.65+-0.17, p<0.0001, resp.). Results are reproducible with coefficient of variation between two duplicate measurements <=8% for all indices. We found that whereas atrial myocardium contains less mitochondria than the ventricle, atrial bioenergetic profiles are comparable to left ventricle. In conclusion, high resolution respirometry has been adapted to homogenates of human cardiac muscle and shown to be reliable and reproducible.
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
30223 - Anaesthesiology
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
2020
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
PLoS One
ISSN
1932-6203
e-ISSN
1932-6203
Volume of the periodical
15
Issue of the periodical within the volume
January
Country of publishing house
US - UNITED STATES
Number of pages
19
Pages from-to
e0226142
UT code for WoS article
000534368000020
EID of the result in the Scopus database
2-s2.0-85077941644