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Mitochondrial redox and pH signaling occurs in axonal and synaptic organelle clusters | Scientific Reports
Description:
Redox switches are important mediators in neoplastic, cardiovascular and neurological disorders. We recently identified spontaneous redox signals in neurons at the single mitochondrion level where transients of glutathione oxidation go along with shortening and re-elongation of the organelle. We now have developed advanced image and signal-processing methods to re-assess and extend previously obtained data. Here we analyze redox and pH signals of entire mitochondrial populations. In total, we quantified the effects of 628 redox and pH events in 1797 mitochondria from intercostal axons and neuromuscular synapses using optical sensors (mito-Grx1-roGFP2; mito-SypHer). We show that neuronal mitochondria can undergo multiple redox cycles exhibiting markedly different signal characteristics compared to single redox events. Redox and pH events occur more often in mitochondrial clusters (medium cluster size: 34.1 ± 4.8 μm2). Local clusters possess higher mitochondrial densities than the rest of the axon, suggesting morphological and functional inter-mitochondrial coupling. We find that cluster formation is redox sensitive and can be blocked by the antioxidant MitoQ. In a nerve crush paradigm, mitochondrial clusters form sequentially adjacent to the lesion site and oxidation spreads between mitochondria. Our methodology combines optical bioenergetics and advanced signal processing and allows quantitative assessment of entire mitochondrial populations.
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nature portfolio privacy policy transgenic thy1-mito-grx1-rogfp2 advertising nature 469 nature 514 nature 508 nature bx51 wide-field microscope thy1-mito-grx1-rogfp2 mice social media provided analytical tools mitochondrial signaling language future research 0/ reprints demonstrate high-frequency content research thiol-based redox switches direct protonation/de-protonation cellular energy supply redox language vivo tissue-wide synchronization oxidative post-translational modifications oxidative stress-induced apoptosis aav-mediated hippocampal expression inter-mitochondrial coupling mechanism pro-oxidative egsh shifts enable high-throughput analysis injected aav1/2-mito-sypher11 custom-written matlab code oscillatory cycle s0 = 4dt multi-parametric imaging techniques aav-mito-sypher explants triangularis sterni explants functional inter-mitochondrial coupling tools smeared high-frequency areas energy metabolism characterize individual grx1-rogfp2 triangularis sterni muscle permissions regulate respiratory bioenergetics vivo imaging reveals stress-induced mechanism ros-induced ros release mitochondrial metabolism triggered glutathione oxidation/ph spikes mitochondrial fluorescence trace accompanying redox/ph fluctuations mito-grx1-rogfp2
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headline:Mitochondrial redox and pH signaling occurs in axonal and synaptic organelle clusters
description:Redox switches are important mediators in neoplastic, cardiovascular and neurological disorders. We recently identified spontaneous redox signals in neurons at the single mitochondrion level where transients of glutathione oxidation go along with shortening and re-elongation of the organelle. We now have developed advanced image and signal-processing methods to re-assess and extend previously obtained data. Here we analyze redox and pH signals of entire mitochondrial populations. In total, we quantified the effects of 628 redox and pH events in 1797 mitochondria from intercostal axons and neuromuscular synapses using optical sensors (mito-Grx1-roGFP2; mito-SypHer). We show that neuronal mitochondria can undergo multiple redox cycles exhibiting markedly different signal characteristics compared to single redox events. Redox and pH events occur more often in mitochondrial clusters (medium cluster size: 34.1â±â4.8âμm2). Local clusters possess higher mitochondrial densities than the rest of the axon, suggesting morphological and functional inter-mitochondrial coupling. We find that cluster formation is redox sensitive and can be blocked by the antioxidant MitoQ. In a nerve crush paradigm, mitochondrial clusters form sequentially adjacent to the lesion site and oxidation spreads between mitochondria. Our methodology combines optical bioenergetics and advanced signal processing and allows quantitative assessment of entire mitochondrial populations.
datePublished:2016-03-22T00:00:00Z
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headline:Mitochondrial redox and pH signaling occurs in axonal and synaptic organelle clusters
description:Redox switches are important mediators in neoplastic, cardiovascular and neurological disorders. We recently identified spontaneous redox signals in neurons at the single mitochondrion level where transients of glutathione oxidation go along with shortening and re-elongation of the organelle. We now have developed advanced image and signal-processing methods to re-assess and extend previously obtained data. Here we analyze redox and pH signals of entire mitochondrial populations. In total, we quantified the effects of 628 redox and pH events in 1797 mitochondria from intercostal axons and neuromuscular synapses using optical sensors (mito-Grx1-roGFP2; mito-SypHer). We show that neuronal mitochondria can undergo multiple redox cycles exhibiting markedly different signal characteristics compared to single redox events. Redox and pH events occur more often in mitochondrial clusters (medium cluster size: 34.1â±â4.8âμm2). Local clusters possess higher mitochondrial densities than the rest of the axon, suggesting morphological and functional inter-mitochondrial coupling. We find that cluster formation is redox sensitive and can be blocked by the antioxidant MitoQ. In a nerve crush paradigm, mitochondrial clusters form sequentially adjacent to the lesion site and oxidation spreads between mitochondria. Our methodology combines optical bioenergetics and advanced signal processing and allows quantitative assessment of entire mitochondrial populations.
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