Isolating the segment of the mitochondrial electron transport chain responsible for mitochondrial damage during cardiac ischemia.
Ischemia damages the mitochondrial electron transport chain (ETC), mediated in part by damage generated by the mitochondria themselves. Mitochondrial damage resulting from ischemia, in turn, leads to cardiac injury during reperfusion. The goal of the present study was to localize the segment of the ETC that produces the ischemic mitochondrial damage. We tested if blockade of the proximal ETC at complex I differed from blockade distal in the chain at cytochrome oxidase. Isolated rabbit hearts were perfused for 15min followed by 30min stop-flow ischemia at 37 degrees C. Amobarbital (2.5mM) or azide (5mM) was used to block proximal (complex I) or distal (cytochrome oxidase) sites in the ETC. Time control hearts were buffer-perfused for 45min. Subsarcolemmal mitochondria (SSM) and interfibrillar mitochondria (IFM) were isolated. Ischemia decreased cytochrome c content in SSM but not in IFM compared to time control. Blockade of electron transport at complex I preserved the cytochrome c content in SSM. In contrast, blockade of electron transport at cytochrome oxidase with azide did not retain cytochrome c in SSM during ischemia. Since blockade of electron transport at complex III also prevented cytochrome c loss during ischemia, the specific site that elicits mitochondrial damage during ischemia is likely located in the segment between complex III and cytochrome oxidase.
Department of Medicine, Division of Cardiology, Case Western Reserve University, Cleveland, OH 44106, United States.
Qun
Q
Chen
Guotian
G
Yin
Sarah
S
Stewart
Ying
Y
Hu
Edward J
EJ
Lesnefsky
D000654:Amobarbital / Q000494:pharmacology; D000818:Animals; D045304:Cytochromes c / Q000378:metabolism; D004195:Disease Models, Animal; D004579:Electron Transport / Q000187:drug effects; D014450:Electron Transport Complex III / Q000378:metabolism*; D003576:Electron Transport Complex IV / Q000378:metabolism*; D066298:In Vitro Techniques; D008929:Mitochondria, Heart / Q000201:enzymology* / Q000473:pathology; D017202:Myocardial Ischemia / Q000201:enzymology* / Q000473:pathology; D011817:Rabbits
D045304:Cytochromes c; D003576:Electron Transport Complex IV; D014450:Electron Transport Complex III; D000654:Amobarbital