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Monitoring Mitochondrial Membranes Permeability in Live Neurons and Mitochondrial Swelling Through Electron Microscopy Analysis

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Neuronal Cell Death

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1254))

Abstract

Maintenance of mitochondrial membrane integrity is essential for mitochondrial function and neuronal viability . Apoptotic stimulus or calcium overload leads to mitochondrial permeability transition pore (mPTP ) opening and induces mitochondrial swelling , a common feature of mitochondrial membrane permeabilization. The first phenomenon can be evaluated in cells loaded with the dye calcein -AM quenched by cobalt , and mitochondrial swelling can be detected by electron microscopy through the analysis of mitochondrial membrane integrity. Here, we describe a live cell imaging assay to detect mitochondrial permeability transition and the development of a detailed analysis of morphological and ultrastructural changes that mitochondria undergo during this process.

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Acknowledgments

This work was supported by grants from the Basal Center of Excellence in Aging and Regeneration (CONICYT—PFB 12/2007) and FONDECYT N° 1120156 to NCI, and a predoctoral fellowship from CONICYT to MSA.

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Authors and Affiliations

  1. Departamento de Biología Celular y Molecular, Facultad de Ciencias Biológicas, Centro de Envejecimiento y Regeneración (CARE), Pontificia Universidad Católica de Chile, Santiago, Chile

    Macarena S. Arrázola & Nibaldo C. Inestrosa

  2. Faculty of Medicine, Center for Healthy Brain Ageing, School of Psychiatry, University of New South Wales, Sydney, Australia

    Nibaldo C. Inestrosa

  3. Pontificia Universidad Católica de Chile, CARE Biomedical Center, Av. Alameda 340, Santiago, Chile

    Nibaldo C. Inestrosa

Authors
  1. Macarena S. Arrázola
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  2. Nibaldo C. Inestrosa
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Corresponding author

Correspondence to Nibaldo C. Inestrosa .

Editor information

Editors and Affiliations

  1. Dept. of Veterinary Sciences, University of Torino, Grugliasco, Torino, Italy

    Laura Lossi

  2. Dept. of Veterinary Sciences, University of Torino, Grugliasco, Torino, Italy

    Adalberto Merighi

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Arrázola, M.S., Inestrosa, N.C. (2015). Monitoring Mitochondrial Membranes Permeability in Live Neurons and Mitochondrial Swelling Through Electron Microscopy Analysis. In: Lossi, L., Merighi, A. (eds) Neuronal Cell Death. Methods in Molecular Biology, vol 1254. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2152-2_7

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  • DOI: https://doi.org/10.1007/978-1-4939-2152-2_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2151-5

  • Online ISBN: 978-1-4939-2152-2

  • eBook Packages: Springer Protocols

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