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Patch-Clamp Recording of the Activity of Ion Channels in the Inner Mitochondrial Membrane

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Mitochondrial Medicine

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2276))

Abstract

Mitochondria are intracellular organelles, which play a crucial role in the generation of ATP. Mitochondria are surrounded by a double membrane, consisting of a smooth outer membrane (OMM) and a markedly folded inner mitochondrial membrane (IMM). Mitochondrion that has been stripped of its outer membrane, leaving the inner membrane intact is called mitoplast. There is a number of different transport proteins located in the inner mitochondrial membrane including ion channels that mediate fluxes of potassium, calcium, and chloride ions. These channels regulate the mitochondrial membrane potential, respiration, and production of reactive oxygen species. The stability of mitoplasts offers the possibility of measuring the activity of ion channels from IMM using the patch-clamp technique. Electrophysiological measurements of currents through ion channels in the IMM permit discovery of unique properties of these channels with the aim of new specific pharmacological therapies. In this chapter, we describe the isolation of mitochondria, preparation of mitoplast for patch-clamp recordings and single-mitoplast PCR experiments, which can be helpful in mastering the technique of recording the activity of mitochondrial ion channels.

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Acknowledgments

This work was supported in part by grants from the National Science Center: 2016/21/B/NZ1/02769 to P.B., 2020/36/T/NZ1/00116 to R.P.K., 2018/31/N/NZ1/00928 to A.W., and 2015/19/B/NZ1/02794 to P.K.

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Authors and Affiliations

  1. Department of Physics and Biophysics, Institute of Biology, Warsaw University of Life Sciences—SGGW, Warsaw, Poland

    Piotr Bednarczyk & Rafał P. Kampa

  2. Laboratory of Intracellular Ion Channels, Nencki Institute of Experimental Biology, Warsaw, Poland

    Rafał P. Kampa, Shur Gałecka, Aleksandra Sęk, Agnieszka Walewska & Piotr Koprowski

  3. Faculty of Chemistry, University of Warsaw, Warsaw, Poland

    Aleksandra Sęk

Authors
  1. Piotr Bednarczyk
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  2. Rafał P. Kampa
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  3. Shur Gałecka
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  4. Aleksandra Sęk
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  5. Agnieszka Walewska
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  6. Piotr Koprowski
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Corresponding author

Correspondence to Piotr Koprowski .

Editor information

Editors and Affiliations

  1. Department of Pharmaceutical Sciences, Midwestern University, Glendale, AZ, USA

    Volkmar Weissig

  2. Cochin Hospital, Cochin Institute, INSERM U1016, PARIS, France

    Marvin Edeas

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Bednarczyk, P., Kampa, R.P., Gałecka, S., Sęk, A., Walewska, A., Koprowski, P. (2021). Patch-Clamp Recording of the Activity of Ion Channels in the Inner Mitochondrial Membrane. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine . Methods in Molecular Biology, vol 2276. Springer, New York, NY. https://doi.org/10.1007/978-1-0716-1266-8_18

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  • DOI: https://doi.org/10.1007/978-1-0716-1266-8_18

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-0716-1265-1

  • Online ISBN: 978-1-0716-1266-8

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