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• W2077284342 abstract "Depolarization of neurons promotes Ca2+ influx through voltage-activated channels, raising the intracellular Ca2+ concentration ([Ca2+]i). The consequences of such changes in [Ca2+]i for mitochondrial function were assessed in single, freshly dissociated mammalian neurons. Microfluorimetric techniques were used to measure [Ca2+]i, mitochondrial membrane potential [delta psi m, Rhodamine 123 (Rh 123) fluorescence], NAD(P)H/NAD(P)+ autofluorescence and flavoprotein autofluorescence combined with whole-cell voltage-clamp techniques. Brief (100-500 ms) depolarization of the cell membrane by high K+ or by voltage commands raised [Ca2+]i and depolarized delta psi m. The change in delta psi m was dependent on extracellular Ca2+. Under voltage-clamp control of the cell membrane, the voltage-dependence of the change in Rh 123 fluorescence reflected that of the Ca2+ current. The response was reduced by Ca2+ buffers introduced into the cell. The behaviour of this signal is thus consistent with a mitochondrial response to raised [Ca2+]i and does not reflect the change in cell membrane potential per se. Similar stimuli caused a rapid decrease of NAD(P)H autofluorescence, followed by an increase which could last several minutes. Flavoprotein fluorescence increased transiently, followed by a decrease lasting for several minutes. These signals indicate an initial oxidation of NAD(P)H and FADH, followed by a prolonged increase in the reduced state of both coenzymes. All these changes were dependent on extracellular [Ca2+]. Raising [Ca2+]i again during the period of NAD+ reduction caused an oxidizing response. Ruthenium Red applied to the cells (i) reduced both the Ca2+ current and the depolarization-induced [Ca2+]i transient and (ii) directly quenched Rh 123 fluorescence. When introduced into the cells with patch pipettes, it prevented the changes in autofluorescence without interfering with the Ca2+ conductance. Oligomycin blocked neither the response of delta psi m nor of NADH autofluorescence, suggesting that the signals do not reflect a response to falling ATP/ADP.Pi ratios as a consequence of the high [Ca2+]i. The changes in NADH autofluorescence were sustained in the presence of iodoacetic acid with pyruvate as substrate. Thus brief physiological elevations of [Ca2+]i depolarize delta psi m, probably through Ca2+ cycling across the mitochondrial inner membrane. The changes in autofluorescence are consistent with (i) increased respiration which could result from the depolarization of delta psi m, followed rapidly by (ii) increased activity of the Ca(2+)-dependent intramitochondrial enzymes. Changes in [Ca2+]i within a physiological range may thus promote significant and long-lasting changes in mitochondrial energy production." @default.
• W2077284342 created "2016-06-24" @default.
• W2077284342 creator A5018410798 @default.
• W2077284342 date "1992-04-01" @default.
• W2077284342 modified "2023-10-12" @default.
• W2077284342 title "Ca2+-dependent changes in the mitochondrial energetics in single dissociated mouse sensory neurons" @default.
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• W2077284342 doi "https://doi.org/10.1042/bj2830041" @default.
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