Mitochondrial and plasma membrane potential of cultured cerebellar neurons during glutamate-induced necrosis, apoptosis, and tolerance

Manus Ward, Heinrich J. Huber, Petronela Weisová, Heiko Düssmann, David G. Nicholls, Jochen H.M. Prehn*

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    98 Scopus citations

    Abstract

    A failure of mitochondrial bioenergetics has been shown to be closely associated with the onset of apoptotic and necrotic neuronal injury. Here, we developed an automated computational model that interprets the single-cell fluorescence for tetramethylrhodamine methyl ester (TMRM) as a consequence of changes in either Δψ m or Δψ p , thus allowing for the characterization of responses for populations of single cells and subsequent statistical analysis. Necrotic injury triggered by prolonged glutamate excitation resulted in a rapid monophasic or biphasic loss of Δψ m that was closely associated with a loss of Δψ p and a rapid decrease in neuronal NADPH and ATP levels. Delayed apoptotic injury, induced by transient glutamate excitation, resulted in a small, reversible decrease in TMRM fluorescence, followed by a sustained hyperpolarization of Δψ m as confirmed using the Δψ p -sensitive anionic probe DiBAC 2 (3). This hyperpolarization of Δψ m was closely associated with a significant increase in neuronal glucose uptake, NADPH availability, and ATP levels. Statistical analysis of the changes in Δψ m or Δψ p at a single-cell level revealed two major correlations; those neurons displaying a more pronounced depolarization of Δψ p during the initial phase of glutamate excitation entered apoptosis more rapidly, and neurons that displayed a more pronounced hyperpolarization of Δψ m after glutamate excitation survived longer. Indeed, those neurons that were tolerant to transient glutamate excitation (18%) showed the most significant increases in Δψ m . Our results indicate that a hyperpolarization of Δψ m is associated with increased glucose uptake, NADPH availability, and survival responses during excitotoxic injury.

    Original languageEnglish (US)
    Pages (from-to)8238-8249
    Number of pages12
    JournalJournal of Neuroscience
    Volume27
    Issue number31
    DOIs
    StatePublished - Aug 1 2007

    Keywords

    • Bioenergetics
    • Excitotoxicity
    • Mitochondria
    • Modeling
    • Necrosis
    • Plasma and mitochondrial membrane potential

    ASJC Scopus subject areas

    • Neuroscience(all)

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