Skip to main content

Mitochondrial Membrane Potential (ΔΨ) Fluctuations Associated with the Metabolic States of Mitochondria

  • Protocol
  • First Online:
Mitochondrial Bioenergetics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1782))

Abstract

The proton electrochemical gradient generated by the respiratory chain activity accounts for over 90% of the available respiratory energy, and, as such, its evaluation and accurate measurement regarding total values and fluctuations are an invaluable component of the understanding of mitochondrial function. Consequently, alterations in electric potential across the inner mitochondrial membrane generated by differential protonic accumulation and transport is known as the mitochondrial membrane potential, or ΔΨ, and is reflective of the functional metabolic status of mitochondria. There are several experimental approaches to measure ΔΨ, ranging from fluorometric evaluations to electrochemical probes. Here, we will expose a particular method for ΔΨ evaluation, which is dependent on the movement of a particular ion, tetraphenylphosphonium (TPP+) with a selective electrode. The evaluation of the accumulation and movements of TPP+ across the inner mitochondrial membrane is a sensitive, immediate, accurate, and simple method of evaluation of ΔΨ in isolated, respiring mitochondria.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Wallace DC (2009) Mitochondria, bioenergetics, and the epigenome in eukaryotic and human evolution. Cold Spring Harb Symp Quant Biol 74:383–393

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Mitchell P (1966) Chemiosmotic coupling in oxidative and photosynthetic phosphorylation. Biol Rev 41:445–501

    Article  CAS  PubMed  Google Scholar 

  3. Jastroch M, Divakaruni AS, Mookerjee S, Treberg JR, Brand MD (2010) Mitochondrial proton and electron leaks. Essays Biochem 47:53–67

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Crofts AR (2004) The Q-cycle – a personal perspective. Photosynth Res 80(1):223–243

    Article  CAS  PubMed  Google Scholar 

  5. Murai M, Miyoshi H (2016) Current topics on inhibitors of respiratory complex I. Biochim Biophys Acta 1857(7):884–891

    Article  CAS  PubMed  Google Scholar 

  6. Azzone GF, Petronilli V, Zoratti M (1984) ‘Cross-talk’ between redox- and ATP-driven H+ pumps. Biochem Soc Trans 12(3):414–416

    Article  CAS  PubMed  Google Scholar 

  7. Gerle C (2016) On the structural possibility of pore-forming mitochondrial FoF1 ATP synthase. Biochim Biophys Acta 1857(8):1191–1196

    Article  CAS  PubMed  Google Scholar 

  8. Brown GC (1992) Control of respiration and ATP synthesis in mammalian mitochondria and cells. Biochem J 284:1–13

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Jensen BD, Gunther TE (1984) The use of tetraphenylphosphonium (TPP±) to measure membrane potentials in mitochondria: membrane binding and respiratory effects. Biophys J 49:105–121

    Google Scholar 

  10. Kamo N, Muratsugu M, Hongoh R, Kobatake Y (1979) Membrane potential of mitochondria measured with an electrode sensitive to tetraphenyl phosphonium and relationship between proton electrochemical potential and phosphorylation potential in steady state. J Membr Biol 49(2):105–121

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

J.S.T. is a recipient of a postdoc scholarship from the Portuguese Fundação para a Ciência e a Tecnologia (SFRH/BPD/94036/2013).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Carlos Marques Palmeira .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Teodoro, J.S., Palmeira, C.M., Rolo, A.P. (2018). Mitochondrial Membrane Potential (ΔΨ) Fluctuations Associated with the Metabolic States of Mitochondria. In: Palmeira, C., Moreno, A. (eds) Mitochondrial Bioenergetics. Methods in Molecular Biology, vol 1782. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7831-1_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7831-1_6

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7830-4

  • Online ISBN: 978-1-4939-7831-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics