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PMID: 11438581 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Spontaneous changes in mitochondrial membrane potential in cultured neurons.

The Journal of neuroscience : the official journal of the Society for Neuroscience ·Vol. 21 ·No. 14 ·2001-07-15 ·Pages 5054-65

Buckman JF, Reynolds IJ

Abstract

Using the mitochondrial membrane potential (DeltaPsi(m))-sensitive fluorescent dyes 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine iodide (JC-1) and tetramethylrhodamine methyl ester (TMRM), we have observed spontaneous changes in the DeltaPsi(m) of cultured forebrain neurons. These fluctuations in DeltaPsi(m) appear to represent partial, transient depolarizations of individual mitochondria. The frequency of these DeltaPsi(m) fluctuations can be significantly lowered by exposure to a photo-induced oxidant burden, an ATP synthase inhibitor, or a glutamate-induced sodium load, without changing overall JC-1 fluorescence intensity. These spontaneous fluctuations in JC-1 signal were not inhibited by altering plasma membrane activity with tetrodotoxin or MK-801 or by blocking the mitochondrial permeability transition pore (PTP) with cyclosporin A. Neurons loaded with TMRM showed similar, low-amplitude, spontaneous fluctuations in DeltaPsi(m). We hypothesize that these DeltaPsi(m) fluctuations are dependent on the proper functioning of the mitochondria and reflect mitochondria alternating between the active and inactive states of oxidative phosphorylation.

MeSH Terms
Animals Benzimidazoles/metabolism Calcium/metabolism Carbocyanines/metabolism Cells, Cultured Chelating Agents/pharmacology Dose-Response Relationship, Drug Egtazic Acid/analogs & derivatives,pharmacology Enzyme Inhibitors/pharmacology Excitatory Amino Acid Agonists/pharmacology Excitatory Amino Acid Antagonists/pharmacology Fluorescent Dyes/metabolism Intracellular Membranes/metabolism Light Membrane Potentials/drug effects,physiology,radiation effects Microscopy, Video Mitochondria/drug effects,metabolism Neurons/cytology,drug effects,metabolism,radiation effects Prosencephalon Rats Rats, Sprague-Dawley Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors Rhodamines/metabolism Tetrodotoxin/pharmacology
Chemicals
Benzimidazoles Carbocyanines Chelating Agents Enzyme Inhibitors Excitatory Amino Acid Agonists Excitatory Amino Acid Antagonists Fluorescent Dyes NMDA receptor A1 Receptors, N-Methyl-D-Aspartate Rhodamines tetramethylrhodamine methyl ester 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine Tetrodotoxin Egtazic Acid Calcium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Buckman J F
Department of Pharmacology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261.
Reynolds I J
References (44)
44 references, click to expand
  1. Physiological cytosolic Ca2+ transients evoke concurrent mitochondrial depolarizations.
    Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12579-83 PMID: 7809081
  2. A test of the singlet oxygen mechanism of cationic dye photosensitization of mitochondrial damage.
    Photochem Photobiol. 1992 Jan;55(1):81-7 PMID: 1603852
  3. Mitochondrial membrane potential and glutamate excitotoxicity in cultured cerebellar granule cells.
    J Neurosci. 2000 Oct 1;20(19):7208-19 PMID: 11007877
  4. Mitochondria and neuronal survival.
    Physiol Rev. 2000 Jan;80(1):315-60 PMID: 10617771
  5. Depolarization of in situ mitochondria due to hydrogen peroxide-induced oxidative stress in nerve terminals: inhibition of alpha-ketoglutarate dehydrogenase.
    J Neurochem. 1999 Jul;73(1):220-8 PMID: 10386974
  6. The calcium response to the excitotoxin kainate is amplified by subsequent reduction of extracellular sodium.
    Neuroscience. 1995 Oct;68(4):1051-7 PMID: 8544981
  7. Mitochondrial dysfunction is a primary event in glutamate neurotoxicity.
    J Neurosci. 1996 Oct 1;16(19):6125-33 PMID: 8815895
  8. Mitochondria, calcium regulation, and acute glutamate excitotoxicity in cultured cerebellar granule cells.
    J Neurochem. 1996 Dec;67(6):2282-91 PMID: 8931459
  9. Mitochondrial membrane potential and neuronal glutamate excitotoxicity: mortality and millivolts.
    Trends Neurosci. 2000 Apr;23(4):166-74 PMID: 10717676
  10. Distinct influx pathways, not calcium load, determine neuronal vulnerability to calcium neurotoxicity.
    J Neurochem. 1998 Dec;71(6):2349-64 PMID: 9832133
  11. Fluctuations in mitochondrial membrane potential caused by repetitive gating of the permeability transition pore.
    Biochem J. 1999 Oct 15;343 Pt 2:311-7 PMID: 10510294
  12. Close contacts with the endoplasmic reticulum as determinants of mitochondrial Ca2+ responses.
    Science. 1998 Jun 12;280(5370):1763-6 PMID: 9624056
  13. The role of intracellular Na+ and mitochondria in buffering of kainate-induced intracellular free Ca2+ changes in rat forebrain neurones.
    J Physiol. 1998 May 15;509 ( Pt 1):103-16 PMID: 9547385
  14. Contributions of mitochondria to animal physiology: from homeostatic sensor to calcium signalling and cell death.
    J Physiol. 1999 Apr 1;516 ( Pt 1):1-17 PMID: 10066918
  15. Mitochondrial transport of cations: channels, exchangers, and permeability transition.
    Physiol Rev. 1999 Oct;79(4):1127-55 PMID: 10508231
  16. Mitochondria as the central control point of apoptosis.
    Trends Cell Biol. 2000 Sep;10(9):369-77 PMID: 10932094
  17. Organelle motility and metabolism in axons vs dendrites of cultured hippocampal neurons.
    J Cell Sci. 1996 May;109 ( Pt 5):971-80 PMID: 8743944
  18. The mitochondrial death/life regulator in apoptosis and necrosis.
    Annu Rev Physiol. 1998;60:619-42 PMID: 9558479
  19. Mitochondria and Na+/Ca2+ exchange buffer glutamate-induced calcium loads in cultured cortical neurons.
    J Neurosci. 1995 Feb;15(2):1318-28 PMID: 7869100
  20. Decoding of cytosolic calcium oscillations in the mitochondria.
    Cell. 1995 Aug 11;82(3):415-24 PMID: 7634331
  21. Mitochondrial and extramitochondrial apoptotic signaling pathways in cerebrocortical neurons.
    Proc Natl Acad Sci U S A. 2000 May 23;97(11):6161-6 PMID: 10811898
  22. Mitochondrial depolarization in glutamate-stimulated neurons: an early signal specific to excitotoxin exposure.
    J Neurosci. 1996 Sep 15;16(18):5688-97 PMID: 8795624
  23. Dynamics, structure, and function are coupled in the mitochondrial matrix.
    Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8057-61 PMID: 1896451
  24. Visualization of cyclosporin A and Ca2+-sensitive cyclical mitochondrial depolarizations in cell culture.
    Biochim Biophys Acta. 1999 Jan 27;1410(1):77-84 PMID: 10076017
  25. The mitochondrial permeability transition.
    Biochim Biophys Acta. 1995 Jul 17;1241(2):139-76 PMID: 7640294
  26. Mitochondria are excitable organelles capable of generating and conveying electrical and calcium signals.
    Cell. 1997 Jun 27;89(7):1145-53 PMID: 9215636
  27. Quantitative evaluation of mitochondrial calcium content in rat cortical neurones following a glutamate stimulus.
    J Physiol. 2001 Mar 15;531(Pt 3):793-805 PMID: 11251059
  28. Microdomains with high Ca2+ close to IP3-sensitive channels that are sensed by neighboring mitochondria.
    Science. 1993 Oct 29;262(5134):744-7 PMID: 8235595
  29. Transient mitochondrial depolarizations reflect focal sarcoplasmic reticular calcium release in single rat cardiomyocytes.
    J Cell Biol. 1998 Aug 24;142(4):975-88 PMID: 9722610
  30. Glutamate-induced neuronal death: a succession of necrosis or apoptosis depending on mitochondrial function.
    Neuron. 1995 Oct;15(4):961-73 PMID: 7576644
  31. Role of calcium ions in regulation of mammalian intramitochondrial metabolism.
    Physiol Rev. 1990 Apr;70(2):391-425 PMID: 2157230
  32. Mitochondria as regulators of apoptosis: doubt no more.
    Biochim Biophys Acta. 1998 Aug 10;1366(1-2):151-65 PMID: 9714783
  33. Inhibition by cyclosporin A of a Ca2+-dependent pore in heart mitochondria activated by inorganic phosphate and oxidative stress.
    Biochem J. 1988 Oct 1;255(1):357-60 PMID: 3196322
  34. The central executioner of apoptosis: multiple connections between protease activation and mitochondria in Fas/APO-1/CD95- and ceramide-induced apoptosis.
    J Exp Med. 1997 Jul 7;186(1):25-37 PMID: 9206994
  35. Glutamate-induced mitochondrial depolarisation and perturbation of calcium homeostasis in cultured rat hippocampal neurones.
    J Physiol. 1999 Sep 1;519 Pt 2:451-66 PMID: 10457062
  36. Mitochondria and apoptosis.
    Science. 1998 Aug 28;281(5381):1309-12 PMID: 9721092
  37. Glutamate-induced neuron death requires mitochondrial calcium uptake.
    Nat Neurosci. 1998 Sep;1(5):366-73 PMID: 10196525
  38. Role of uncoupled and non-coupled oxidations in maintenance of safely low levels of oxygen and its one-electron reductants.
    Q Rev Biophys. 1996 May;29(2):169-202 PMID: 8870073
  39. Cyclosporin A delays mitochondrial depolarization induced by N-methyl-D-aspartate in cortical neurons: evidence of the mitochondrial permeability transition.
    Neuroscience. 1996 Dec;75(4):993-7 PMID: 8938735
  40. Mitochondria and programmed cell death: back to the future.
    FEBS Lett. 1996 Oct 28;396(1):7-13 PMID: 8906857
  41. Parallel oscillations of intracellular calcium activity and mitochondrial membrane potential in mouse pancreatic B-cells.
    Biochem Biophys Res Commun. 2000 Jan 7;267(1):179-83 PMID: 10623595
  42. Mitochondria accumulate Ca2+ following intense glutamate stimulation of cultured rat forebrain neurones.
    J Physiol. 1997 Jan 1;498 ( Pt 1):31-47 PMID: 9023766
  43. Energy transduction in intact synaptosomes. Influence of plasma-membrane depolarization on the respiration and membrane potential of internal mitochondria determined in situ.
    Biochem J. 1980 Jan 15;186(1):21-33 PMID: 7370008
  44. Effects of oxidants and glutamate receptor activation on mitochondrial membrane potential in rat forebrain neurons.
    J Neurochem. 1998 Dec;71(6):2392-400 PMID: 9832137
Article Info
Journal
The Journal of neuroscience : the official journal of the Society for Neuroscience
Abbr.
J Neurosci
ISSN
1529-2401
Published
2001-07-15
Pages
5054-65
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC6762873
Subset
IM
Grants
NINDS NIH HHS · F32NS11147 · United States
NINDS NIH HHS · T32NS07391 · United States
External Links
PubMed source PMC Full Text View journal details
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