scholarly journals Mitochondria Exert a Negative Feedback on the Propagation of Intracellular Ca2+ Waves in Rat Cortical Astrocytes

1999 ◽  
Vol 145 (4) ◽  
pp. 795-808 ◽  
Author(s):  
Eric Boitier ◽  
Ruth Rea ◽  
Michael R. Duchen
Keyword(s):  
Mitochondrial Membrane ◽  
Imaging Techniques ◽  
Mitochondrial Depolarization ◽  
Cortical Astrocytes ◽  
Propagating Waves ◽  
Carbonyl Cyanide ◽  
Rate Of Decay ◽  
Digital Fluorescence Imaging ◽  
Mitochondrial Uncoupler ◽  
Cell Collapse

We have used digital fluorescence imaging techniques to explore the interplay between mitochondrial Ca2+ uptake and physiological Ca2+ signaling in rat cortical astrocytes. A rise in cytosolic Ca2+ ([Ca2+]cyt), resulting from mobilization of ER Ca2+ stores was followed by a rise in mitochondrial Ca2+ ([Ca2+]m, monitored using rhod-2). Whereas [Ca2+]cyt recovered within ∼1 min, the time to recovery for [Ca2+]m was ∼30 min. Dissipating the mitochondrial membrane potential (Δψm, using the mitochondrial uncoupler carbonyl cyanide p-trifluoromethoxy-phenyl-hydrazone [FCCP] with oligomycin) prevented mitochondrial Ca2+ uptake and slowed the rate of decay of [Ca2+]cyt transients, suggesting that mitochondrial Ca2+ uptake plays a significant role in the clearance of physiological [Ca2+]cyt loads in astrocytes. Ca2+ signals in these cells initiated either by receptor-mediated ER Ca2+ release or mechanical stimulation often consisted of propagating waves (measured using fluo-3). In response to either stimulus, the wave traveled at a mean speed of 22.9 ± 11.2 μm/s (n = 262). This was followed by a wave of mitochondrial depolarization (measured using tetramethylrhodamine ethyl ester [TMRE]), consistent with Ca2+ uptake into mitochondria as the Ca2+ wave traveled across the cell. Collapse of Δψm to prevent mitochondrial Ca2+ uptake significantly increased the rate of propagation of the Ca2+ waves by 50%. Taken together, these data suggest that cytosolic Ca2+ buffering by mitochondria provides a potent mechanism to regulate the localized spread of astrocytic Ca2+ signals.

Role of sodium-calcium exchange in regulation of intracellular calcium in nerve terminals

1987 ◽  
Vol 252 (6) ◽  
pp. C595-C603 ◽  
Author(s):  
S. Sanchez-Armass ◽  
M. P. Blaustein
Keyword(s):  
Hill Coefficient ◽  
Nerve Terminals ◽  
Physiological Load ◽  
Carbonyl Cyanide ◽  
Ca Uptake ◽  
Na Ions ◽  
Presynaptic Nerve Terminals ◽  
45Ca Efflux ◽  
Mitochondrial Uncoupler

Ca efflux from rat brain presynaptic nerve terminals (synaptosomes) was examined after loading the terminals with 45Ca during a brief depolarization, usually in media containing 20 microM Ca labeled with 45Ca, to assure a small (physiological) load. Efflux of 45Ca was very slow in the absence of external Na and Ca (approximately 0.5% of the load/s) and was greatly accelerated by Na and/or Ca (presumably Na+-Ca2+ and Ca2+-Ca2+ exchange, respectively). The dependence of 45Ca efflux on external Na was sigmoid, with a Hill coefficient of approximately 2.5; this implies that more than two external Na ions are required to activate the efflux of one Ca ion. The external Na (Nao)-dependent Ca efflux was inhibited by 1 mM external La, by low temperature (Q10 congruent to 2.3), and by raising external K (to depolarize the synaptosomes). With small Ca loads, the mitochondrial uncoupler, carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), had negligible effect on either Ca uptake or efflux; with large loads (greater than or equal to 5 nmol/mg protein), however, FCCP reduced the depolarization-stimulated Ca uptake and increased the Nao-dependent Ca efflux. These effects may be attributed to reduction of mitochondrial Ca sequestration. Mitochondria do not appear to sequester much Ca when the loads are smaller (and more physiological). Estimations of Ca efflux indicate that approximately 20% of a small 45Ca load (approximately 0.75 nmol Ca/mg protein) may be extruded via Na+-Ca2+ exchange within 1 s; this corresponds to a net Ca efflux of approximately 110 pmol Ca X mg protein-1 X s-1.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Genetic Mutations and Mitochondrial Toxins Shed New Light on the Pathogenesis of Parkinson's Disease

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Shigeto Sato ◽  
Nobutaka Hattori
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Mitochondrial Dysfunction ◽  
Cultured Cells ◽  
Cellular Functions ◽  
Carbonyl Cyanide ◽  
Mitochondrial Toxins ◽  
Cellular Abnormalities ◽  
Mitochondrial Uncoupler

The cellular abnormalities in Parkinson's disease (PD) include mitochondrial dysfunction and oxidative damage, which are probably induced by both genetic predisposition and environmental factors. Mitochondrial dysfunction has long been implicated in the pathogenesis of PD. The recent discovery of genes associated with the etiology of familial PD has emphasized the role of mitochondrial dysfunction in PD. The discovery and increasing knowledge of the function of PINK1 and parkin, which are associated with the mitochondria, have also enhanced the understanding of cellular functions. The PINK1-parkin pathway is associated with quality control of the mitochondria, as determined in cultured cells treated with the mitochondrial uncoupler carbonyl cyanide m-chlorophenylhydrazone (CCCP), which causes mitochondrial depolarization. To date, the use of mitochondrial toxins, for example, 1-methyl-4-phynyl-tetrahydropyridine (MPTP) and CCCP, has contributed to our understanding of PD. We review how these toxins and familial PD gene products are associated with and have enhanced our understanding of the role of mitochondrial dysfunction in PD.

Depolarized Mitochondrial Membrane Potential and Protection with Duroquinone in Isolated Perfused Lungs from Rats Exposed to Hyperoxia

2021 ◽  
Author(s):  
Said H. Audi ◽  
Swetha Ganesh ◽  
Pardis Taheri ◽  
Xiao Zhang ◽  
Ranjan K. Dash ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Complex I ◽  
Mitochondrial Membrane ◽  
Rhodamine 6G ◽  
Computational Approach ◽  
Experimental Protocol ◽  
Perfusion System ◽  
The Impact ◽  
Mitochondrial Uncoupler

Dissipation of mitochondrial membrane potential (Δψm) is a hallmark of mitochondrial dysfunction. our objective was to use a previously developed experimental-computational approach to estimate tissue Δψm in intact lungs of rats exposed to hyperoxia, and to evaluate the ability of duroquinone (DQ) to reverse any hyperoxia-induced depolarization of lung Δψm. Rats were exposed to hyperoxia (>95% O2) or normoxia (room air) for 48 hrs, after which lungs were excised and connected to a ventilation-perfusion system. The experimental protocol consisted of measuring the concentration of the fluorescent dye rhodamine 6G (R6G) during three single-pass phases: loading, washing, and uncoupling, in which the lungs were perfused with and without R6G, and with the mitochondrial uncoupler FCCP, respectively. For normoxic lungs, the protocol was repeated with 1) rotenone (complex I inhibitor), 2) rotenone and either DQ or its vehicle (DMSO), and 3) rotenone, glutathione (GSH), and either DQ or DMSO added to the perfusate. Hyperoxic lungs were studied with and without DQ and GSH added to the perfusate. Computational modeling was used to estimate lung Δψm from R6G data. Rat exposure to hyperoxia resulted in partial depolarization (-33 mV) of lung Δψm, and complex I inhibition depolarized lung Δψm by -83 mV. Results also demonstrate the efficacy of DQ to fully reverse both rotenone-induced and hyperoxia-induced lung Δψm depolarization. This study demonstrates hyperoxia-induced Δψm depolarization in intact lungs, and the utility of this approach for assessing the impact of potential therapies such as exogenous quinones that target mitochondria in intact lungs.

Sub-basalt Marchenko imaging with offshore Brazil field data

Geophysics ◽  
2021 ◽  
pp. 1-47
Author(s):  
Xueyi Jia ◽  
Anatoly Baumstein ◽  
Charlie Jing ◽  
Erik Neumann ◽  
Roel Snieder
Keyword(s):  
Seismic Waves ◽  
Mode Conversion ◽  
Imaging Techniques ◽  
Hydrocarbon Exploration ◽  
Sampled Data ◽  
Geological Interpretation ◽  
Propagating Waves ◽  
Seismic Acquisition ◽  
Internal Multiples ◽  
Multiple Elimination

Sub-basalt imaging for hydrocarbon exploration faces challenges with the presence of multiple scattering, attenuation and mode-conversion as seismic waves encounter highly heterogeneous and rugose basalt layers. A combination of modern seismic acquisition that can record densely-sampled data, and advanced imaging techniques make imaging through basalt feasible. Yet, the internal multiples, if not properly handled during seismic processing, can be mapped to reservoir layers by conventional imaging methods, misguiding geological interpretation. Traditional internal multiple elimination methods suffer from the requirement of picking horizons of multiple generators and/or a top-down adaptive subtraction process. Marchenko imaging provides an alternative solution to directly remove the artifacts due to internal multiples, without the need of horizon picking or subtraction. In this paper, we present a successful application of direct Marchenko imaging for sub-basalt de-multiple and imaging with an offshore Brazil field dataset. The internal multiples in this example are generated from the seabed and basalt layers, causing severe artifacts in conventional seismic images. We demonstrate that these artifacts are largely suppressed with Marchenko imaging and propose a general work flow for data pre-processing and regularization of marine streamer datasets. We show that horizontally propagating waves can also be reconstructed by the Marchenko method at far offsets.

Monitoring of mitochondrial membrane potential in isolated perfused rat heart

1984 ◽  
Vol 247 (4) ◽  
pp. H508-H516
Author(s):  
R. A. Kauppinen ◽  
I. E. Hassinen
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Optical Methods ◽  
Perfused Heart ◽  
Rat Hearts ◽  
Carbonyl Cyanide ◽  
Perfused Rat Hearts ◽  
Beating Rate ◽  
Isolated Perfused Rat Hearts

Optical methods were tested for measuring the membrane potential changes of mitochondria in isolated perfused rat hearts. Safranin was found to be rapidly taken up by the Langendorff-perfused heart, and after loading with the dye there was practically no washout of the stain during perfusion with Krebs-Ringer bicarbonate solution. Staining with safranin induced the appearance of an intense absorption band in the reflectance spectrum of the heart, but the absorbance spectrum changes were not useful for monitoring the mitochondrial membrane potential changes because of interference by endogenous hemoproteins. The fluorescence intensity, however, responded in a manner which indicated that its changes originated from dye attached to the mitochondria. A decrease of the fluorescence was found on energizing the mitochondria by decreasing the cellular energy consumption by arrest induced by 18 mM K+ or by decreasing the beating rate of an electrically paced heart from 5 Hz to the endogenous ventricular frequency of 1.5 Hz. In hearts arrested by Ca2+ depletion, 18 mM K+ did not affect the safranin fluorescence. This was taken to indicate that under these conditions the safranin fluorescence was not sensitive to the plasma membrane potential. The uncoupler carbonyl cyanide m-chlorophenylhydrazone induced an intense enhancement of safranin fluorescence in the perfused heart, demonstrating that the probe is sensitive to mitochondrial membrane potential.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of Mitochondrion on Calcium Transients at Intact Presynaptic Terminals Depend on Frequency of Nerve Firing

1998 ◽  
Vol 80 (1) ◽  
pp. 186-195 ◽  
Author(s):  
Yan-Yi Peng
Keyword(s):  
Firing Frequency ◽  
Sympathetic Ganglia ◽  
Calcium Transients ◽  
Repetitive Firing ◽  
Rapid Rate ◽  
Presynaptic Terminals ◽  
Carbonyl Cyanide ◽  
Frequency Of Stimulation ◽  
Mitochondrial Uncoupler ◽  
Similar Peak

Peng, Yan-Yi. Effects of mitochondrion on calcium transients at intact presynaptic terminals depend on frequency of nerve firing. J Neurophysiol. 80: 186–195, 1998. The rate and the total amount of Ca2+ elevation in the presynaptic terminals of bullfrog sympathetic ganglia depend on the firing frequency of the terminals. Carbonyl cyanide m-chlorophenylhydrazone (CCCP), a mitochondrial uncoupler, was used for testing whether mitochondrial Ca2+ uptake is one of the mechanisms that underlie this frequency dependence. Fura-2 fluorimetry was used for measurement of intraterminal Ca2+. When stimulations of different durations (30 and 1.5 s) and frequencies (4 and 20 Hz) evoked Ca2+ transients with similar peak amplitudes (264 ± 22 nM vs. 251 ± 18 nM, means ± SE), CCCP augmented the responses to the 4-Hz stimulation 8.9 times more strongly than it did the responses to the 20-Hz stimulation (249.7 ± 81.5% vs. 25.3 ± 10.2%). When stimulations delivered at the two frequencies had the same durations (1.5, 3, 6, 10, 20, and 30 s), CCCP enlarged the responses to the 4-Hz stimulations up to 4.2 times more than it did the responses to the 20-Hz stimulations. When the same number of stimuli (120) was delivered at the two frequencies, the effects of CCCP on the responses evoked by the 4-Hz train were again 6.8 times stronger than its effects on the responses to the 20-Hz stimulation. Therefore neither the peak amplitudes of the responses nor the durations of the stimulations dictated the extent to which the mitochondria modulated the peak [Ca2+]i. Instead, the extent of the modulation was governed by the frequency of stimulation. Specifically, the less frequent the Ca2+ influx, the stronger the mitochondrial modulation. Also, during nerve firing Ca2+ release from the ryanodine-sensitive store had a higher potential to influence the [Ca2+]i transients than did Ca2+ removal by the mitochondria for the first 6 s of the responses. On cessation of stimulation, CCCP reduced the initial rapid rate of Ca2+ decay. Thus uptake by the mitochondria was an important mechanism for Ca2+ removal after repetitive firing at the presynaptic terminals.

scholarly journals Magnesium content and net fluxes in squid giant axons.

1979 ◽  
Vol 74 (6) ◽  
pp. 739-752 ◽  
Author(s):  
M Caldwell-Violich ◽  
J Requena
Keyword(s):  
Atomic Absorption ◽  
Atomic Absorption Spectroscopy ◽  
Magnesium Content ◽  
Iodoacetic Acid ◽  
Half Time ◽  
Electrochemical Gradient ◽  
Carbonyl Cyanide ◽  
Mg Content ◽  
Mitochondrial Uncoupler ◽  
Net Fluxes

The Mg content of axons freshly dissected from living specimens of the tropical squid Doryteuthis plei was determined by atomic absorption spectroscopy to be 4.2 +/- 0.2 mmol/kg axoplasm. The axon's ability to maintain this physiological content of total intracellular Mg([Mg]i) was studied. Mgi was shown to be a linear function of Mgo when Mgo of incubating fluid was varied between 0 and 250 mM. When Mgo = 15 mM, Mgi was found to be the same in incubated fibers as in fibers freshly dissected. Mgi levels were unaffected by depolarization of the membrane by high Ko. Stimulation resulted in an extra influx of Mg of 0.05 pmol/(cm2 . impulse) when Mgo = 55 mM. Mgi was found to be a complicated function of the concentration of extracellular Na or Li (Xo), which was substituted for Tris. With 385 mM Lio the Mgi level was found to be 2.5-fold larger than the level observed with 385 mM Nao after incubation for 3 h. The function relating Mgo to Xo was qualitatively unaffected in axons poisoned with the mitochondrial uncoupler carbonyl cyanide, p-trifluorome-thoxy-phenylhydrazone (FCCP) and the inhibitor of glycolysis, iodoacetic acid (IAA); the absolute levels of Mgi, however, were some 30% higher in the poisoned axons at all [X]o explored. 2 h incubation of axons in a 333 mM Mg, 40 mM Li solution increased Mgi 3.5-fold in control axons and 5-fold in poisoned axons. These Mg-loaded axons were able to recover physiological levels of Mgi with a half-time of 3-5 h only if kept in a solution which contained Na (220 mM) regardless of whether the axons had been inhibited with FCCP + IAA. Therefore, it may be concluded that the physiological Mgi concentration can be maintained by the Na electrochemical gradient, even when the axon is metabolically poisoned.

scholarly journals Mitochondrial depolarization and electrophysiological changes during ischemia in the rabbit and human heart

2014 ◽  
Vol 307 (8) ◽  
pp. H1178-H1186 ◽  
Author(s):  
Matthew S. Sulkin ◽  
Bas J. Boukens ◽  
Megan Tetlow ◽  
Sarah R. Gutbrod ◽  
Fu Siong Ng ◽  
...  
Keyword(s):  
Mitochondrial Membrane ◽  
Human Heart ◽  
Transmembrane Potential ◽  
Optical Mapping ◽  
Imaging Modality ◽  
Ischemia Reperfusion ◽  
Mitochondrial Depolarization ◽  
Inner Mitochondrial Membrane ◽  
Ventricular Tissue ◽  
Direct Association

Instability of the inner mitochondrial membrane potential (ΔΨm) has been implicated in electrical dysfunction, including arrhythmogenesis during ischemia-reperfusion. Monitoring ΔΨm has led to conflicting results, where depolarization has been reported as sporadic and as a propagating wave. The present study was designed to resolve the aforementioned difference and determine the unknown relationship between ΔΨm and electrophysiology. We developed a novel imaging modality for simultaneous optical mapping of ΔΨm and transmembrane potential ( Vm). Optical mapping was performed using potentiometric dyes on preparations from 4 mouse hearts, 14 rabbit hearts, and 7 human hearts. Our data showed that during ischemia, ΔΨm depolarization is sporadic and changes asynchronously with electrophysiological changes. Spatially, ΔΨm depolarization was associated with action potential duration shortening but not conduction slowing. Analysis of focal activity indicated that ΔΨm is not different within the myocardium where the focus originates compared with normal ventricular tissue. Overall, our data suggest that during ischemia, mitochondria maintain their function at the expense of sarcolemmal electrophysiology, but ΔΨm depolarization does not have a direct association to ischemia-induced arrhythmias.

scholarly journals The oxidation of glutamine and glutamate in relation to anion transport in enterocyte mitochondria

1984 ◽  
Vol 218 (2) ◽  
pp. 449-458 ◽  
Author(s):  
D F Evered ◽  
B Masola
Keyword(s):  
Amino Acids ◽  
Potassium Phosphate ◽  
Potassium Acetate ◽  
Anion Transport ◽  
Mitochondrial Swelling ◽  
O2 Uptake ◽  
Carbonyl Cyanide ◽  
Mitochondrial Uncoupler ◽  
Main Substrate ◽  
Stimulation Of

The oxidation of L-glutamate and L-glutamine by enterocyte mitochondria was supported by malate. The stimulation of the rate of oxidation of the two amino acids by small amounts of added malate was 93% and 76% respectively. This could not be accounted for by the oxidation of the small amounts of malate added. Amino-oxyacetate added initially inhibited malate-supported oxidation of L-glutamate by 81% and that of L-glutamine by 38%. The inhibition of L-glutamate oxidation was partially reversed by L-glutamine. The dicarboxylate-carrier inhibitor 2-phenylsuccinate inhibited the malate-supported oxidation of both amino acids, but appeared to be slightly stimulatory to L-glutamine oxidation when added initially. The inhibition of L-glutamate oxidation was reversed by L-glutamine. The mitochondrial uncoupler FCCP (carbonyl cyanide p-trifluoromethoxyphenylhydrazone) inhibited malate-supported oxidation of L-glutamate by 78% when added initially. The oxidation of L-glutamine was completely inhibited. However, the uncoupler stimulated the oxidation of both amino acids when added finally. Pyruvate inhibited aspartate synthesis when either of these amino acids was the main substrate, alanine being synthesized. There was no effect on O2 uptake. Mitochondria did not swell in KCl solution, but swelled rapidly in water. Mitochondrial swelling in potassium phosphate and potassium acetate solutions was activated by valinomycin and to a lesser extent by the further addition of FCCP. With potassium malate, swelling was mainly activated by phosphate. The swelling of enterocyte mitochondria in potassium glutamate was slow. In glutamine solution, mitochondrial swelling was greater and appeared to be enhanced by the initial presence of small amounts of phosphate.

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