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The Journal of Neuroscience, July 15, 2001, 21(14):5054-5065
Spontaneous Changes in Mitochondrial Membrane Potential in
Cultured Neurons
Jennifer F.
Buckman and
Ian J.
Reynolds
Department of Pharmacology, University of Pittsburgh, Pittsburgh,
Pennsylvania 15261
Using the mitochondrial membrane potential
(  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 m of cultured
forebrain neurons. These fluctuations in m appear to
represent partial, transient depolarizations of individual
mitochondria. The frequency of these 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 m. We hypothesize that these m
fluctuations are dependent on the proper functioning of the
mitochondria and reflect mitochondria alternating between the active
and inactive states of oxidative phosphorylation.
Key words:
mitochondria; membrane potential; ATP; JC-1; TMRM; F1FO ATPase
Copyright © 2001 Society for Neuroscience 0270-6474/01/21145054-12$05.00/0
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