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Property "Has-20abstract" (as page type) with input value "Mitochondrial-20reactive-20oxygen-20species-20(ROS)-20are-20important-20cellular-20signaling-20molecules,-20but-20can-20cause-20oxidative-20damage-20if-20not-20kept-20within-20tolerable-20limits.-20An-20important-20proximal-20form-20of-20ROS-20in-20mitochondria-20is-20superoxide.-20Its-20production-20is-20thought-20to-20occur-20in-20regulated-20stochastic-20bursts,-20but-20current-20methods-20using-20mitochondrial-20targeted-20cpYFP-20to-20assess-20superoxide-20flashes-20are-20confounded-20by-20changes-20in-20pH.-20Accordingly,-20these-20flashes-20are-20generally-20referred-20to-20as-20'mitoflashes'.-20Here-20we-20provide-20regulatory-20insights-20into-20mitoflashes-20and-20pH-20fluctuations-20in-20skeletal-20muscle,-20and-20the-20role-20of-20uncoupling-20protein-2D3-20(UCP3).-20Using-20quantitative-20confocal-20microscopy-20of-20mitoflashes-20in-20intact-20muscle-20fibers,-20we-20show-20that-20the-20mitoflash-20magnitude-20significantly-20correlates-20with-20the-20degree-20of-20mitochondrial-20inner-20membrane-20depolarization-20and-20ablation-20of-20UCP3-20did-20not-20affect-20this-20correlation.-20We-20assessed-20the-20effects-20of-20the-20absence-20of-20UCP3-20on-20mitoflash-20activity-20in-20intact-20skeletal-20muscle-20fibers,-20and-20found-20no-20effects-20on-20mitoflash-20frequency,-20amplitude-20or-20duration,-20with-20a-20slight-20reduction-20in-20the-20average-20size-20of-20mitoflashes.-20We-20further-20investigated-20the-20regulation-20of-20pH-20flashes-20(pHlashes,-20presumably-20a-20component-20of-20mitoflash)-20by-20UCP3-20using-20mitochondrial-20targeted-20SypHer-20(mt-2DSypHer)-20in-20skeletal-20muscle-20fibers.-20The-20frequency-20of-20pHlashes-20was-20significantly-20reduced-20in-20the-20absence-20of-20UCP3,-20without-20changes-20in-20other-20flash-20properties.-20ROS-20scavenger,-20tiron,-20did-20not-20alter-20pHlash-20frequency-20in-20either-20WT-20or-20UCP3KO-20mice.-20High-20resolution-20respirometry-20revealed-20that-20in-20the-20absence-20of-20UCP3-20there-20is-20impaired-20proton-20leak-20and-20Complex-20I-2Ddriven-20respiration-20and-20maximal-20coupled-20respiration.-20Total-20cellular-20production-20of-20hydrogen-20peroxide-20(H2O2)-20as-20detected-20by-20Amplex-2DUltraRed-20was-20unaffected.-20Altogether,-20we-20demonstrate-20a-20correlation-20between-20mitochondrial-20membrane-20potential-20and-20mitoflash-20magnitude-20in-20skeletal-20muscle-20fibers-20that-20is-20independent-20of-20UCP3-20and-20a-20role-20for-20UCP3-20in-20the-20control-20of-20pHlash-20frequency-20and-20of-20proton-20leak-2D-20and-20Complex-20I-20coupled-2Drespiration-20in-20skeletal-20muscle-20fibers.-20The-20differential-20regulation-20of-20mitoflashes-20and-20pHlashes-20by-20UCP3-20and-20tiron-20also-20indicate-20that-20the-20two-20events,-20though-20may-20be-20related,-20are-20not-20identical-20events.-0A-0A-3Csmall-3ECopyright-20©-202019.-20Published-20by-20Elsevier-20Inc.-3C-2Fsmall-3E" contains invalid characters or is incomplete and therefore can cause unexpected results during a query or annotation process.

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