{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T16:35:26Z","timestamp":1782923726739,"version":"3.54.5"},"reference-count":19,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,4,12]],"date-time":"2021-04-12T00:00:00Z","timestamp":1618185600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Med."],"abstract":"<jats:p><jats:bold>Background:<\/jats:bold> Identification of the mechanisms underlying mitochondrial dysfunction is key to understanding the pathophysiology of acute injuries such as cardiac arrest (CA); however, effective methods for measurement of mitochondrial function associated with mitochondrial isolation have been debated for a long time. This study aimed to evaluate the dysregulation of mitochondrial respiratory function after CA while testing the sampling bias that might be induced by the mitochondrial isolation method.<\/jats:p><jats:p><jats:bold>Materials and Methods:<\/jats:bold> Adult rats were subjected to 10-min asphyxia-induced CA. 30 min after resuscitation, the brain and kidney mitochondria from animals in sham and CA groups were isolated (<jats:italic>n<\/jats:italic> = 8, each). The mitochondrial quantity, expressed as protein concentration (isolation yields), was determined, and the oxygen consumption rates were measured. ADP-dependent (state-3) and ADP-limited (state-4) respiration activities were compared between the groups. Mitochondrial quantity was evaluated based on citrate synthase (CS) activity and cytochrome c concentration, measured independent of the isolation yields.<\/jats:p><jats:p><jats:bold>Results:<\/jats:bold> The state-3 respiration activity and isolation yield in the CA group were significantly lower than those in the sham group (brain, <jats:italic>p<\/jats:italic> &amp;lt; 0.01; kidney, <jats:italic>p<\/jats:italic> &amp;lt; 0.001). The CS activity was significantly lower in the CA group as compared to that in the sham group (brain, <jats:italic>p<\/jats:italic> &amp;lt; 0.01; kidney, <jats:italic>p<\/jats:italic> &amp;lt; 0.01). Cytochrome c levels in the CA group showed a similar trend (brain, <jats:italic>p<\/jats:italic> = 0.08; kidney, <jats:italic>p<\/jats:italic> = 0.25).<\/jats:p><jats:p><jats:bold>Conclusions:<\/jats:bold> CA decreased mitochondrial respiration activity and the quantity of mitochondria isolated from the tissues. Owing to the nature of fragmented or damaged mitochondrial membranes caused by acute injury, there is a potential loss of disrupted mitochondria. Thus, it is plausible that the mitochondrial function in the acute-injury model may be underestimated as this loss is not considered.<\/jats:p>","DOI":"10.3389\/fmed.2021.666735","type":"journal-article","created":{"date-parts":[[2021,4,12]],"date-time":"2021-04-12T04:56:28Z","timestamp":1618203388000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["Methodological Issue of Mitochondrial Isolation in Acute-Injury Rat Model: Asphyxia Cardiac Arrest and Resuscitation"],"prefix":"10.3389","volume":"8","author":[{"given":"Tomoaki","family":"Aoki","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Okuma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lance 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