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Monitoring Mitophagy in Neuronal Cell Cultures

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Neurodegeneration

Part of the book series: Methods in Molecular Biology ((MIMB,volume 793))

Abstract

Proper control of mitochondrial turnover is critical for maintenance of cellular energetics under basal and stressed conditions, and for prevention of endogenous oxidative stress. Whole organelle turnover is mediated through macroautophagy, a process by which autophagosomes deliver mitochondria to the lysosome for hydrolytic degradation. While mitochondrial autophagy can occur as part of a nonselective upregulation of autophagy, selective degradation of damaged or unneeded mitochondria (mitophagy) is a rapidly growing area in development, cancer, and neurodegeneration, particularly with regard to Parkinson’s disease. Due to its dynamic nature, and the potential for regulatory perturbation by disease processes, no single technique is sufficient to evaluate mitophagy. Here, we describe several complementary techniques that include electron microscopy, single cell analysis of LC3 fluorescent puncta, and Western blot, each used in conjunction with a flux inhibitor to trap newly formed autophagosomes in order to monitor mitophagy in neuronal cells.

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Acknowledgments

This work was supported in part by the National Institutes of Health (AG026389, NS065789). CTC is recipient of an AFAR/Ellison Medical Foundation Julie Martin Mid-Career Award in Aging Research.

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Authors and Affiliations

  1. Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    Jianhui Zhu, Ruben K. Dagda & Charleen T. Chu

Authors
  1. Jianhui Zhu
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  2. Ruben K. Dagda
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  3. Charleen T. Chu
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Corresponding author

Correspondence to Charleen T. Chu .

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Editors and Affiliations

  1. Weill Medical College, Dept. Neurology & Neuroscience, Cornell University, East 68th St. 525, New York, 10065, New York, USA

    Giovanni Manfredi

  2. York Avenue 1300, New York, 10065, New York, USA

    Hibiki Kawamata

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Zhu, J., Dagda, R.K., Chu, C.T. (2011). Monitoring Mitophagy in Neuronal Cell Cultures. In: Manfredi, G., Kawamata, H. (eds) Neurodegeneration. Methods in Molecular Biology, vol 793. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-328-8_21

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  • DOI: https://doi.org/10.1007/978-1-61779-328-8_21

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-327-1

  • Online ISBN: 978-1-61779-328-8

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