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Related Experiment Videos

Spontaneous changes in mitochondrial membrane potential in single isolated brain mitochondria.

Olga Vergun1, Tatyana V Votyakova, Ian J Reynolds

  • 1Department of Pharmacology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.

Biophysical Journal
|October 29, 2003
PubMed
Summary

Mitochondrial membrane potential (DeltaPsim) fluctuates spontaneously in about 70% of isolated brain mitochondria, indicating an unstable state. These fluctuations may signal or lead to mitochondrial dysfunction, distinct from permeability transition.

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Area of Science:

  • Mitochondrial physiology
  • Cellular bioenergetics

Background:

  • Mitochondrial membrane potential (DeltaPsim) is crucial for cellular energy production.
  • Spontaneous fluctuations in DeltaPsim have been observed in intact cells but not well characterized in isolated mitochondria.

Purpose of the Study:

  • To investigate spontaneous DeltaPsim fluctuations in single isolated brain mitochondria.
  • To characterize the behavior of fluctuating versus stable mitochondria.

Main Methods:

  • Measurement of DeltaPsim in single isolated brain mitochondria using rhodamine 123 fluorescence.
  • Superfusion of mitochondria with a K(+)-based HEPES-buffer medium containing malate and glutamate.
  • Application of calcium and substrate removal to assess mitochondrial response.

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Main Results:

  • Approximately 70% of energized mitochondria exhibited large amplitude spontaneous DeltaPsim fluctuations.
  • Stable mitochondria either remained stable or transitioned to fluctuating, while fluctuating mitochondria did not stabilize.
  • Fluctuating mitochondria responded rapidly to substrate removal or calcium, unlike stable mitochondria which depolarized with a delay.
  • Fluctuations were independent of oxidant production and permeability transition.

Conclusions:

  • Spontaneous DeltaPsim fluctuations represent an intermediate, unstable mitochondrial state.
  • This unstable state may precede or indicate mitochondrial dysfunction.
  • Isolated mitochondria can exhibit complex dynamic behaviors relevant to cellular health.