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Measuring the Impact of Bile Acids on the Membrane Order of Primary Hepatocytes and Isolated Mitochondria by Fluorescence Imaging and Spectroscopy

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Experimental Cholestasis Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1981))

Abstract

Cholestasis is characterized by impaired secretion of bile flow that can result in the accumulation of highly abnormal levels of bile acids causing hepatocyte and biliary injury. As amphipathic molecules, bile acids can intercalate in lipid membranes, and pathophysiologic concentrations of bile acids have the potential to induce marked changes in the biophysical properties of biomembranes, including membrane ordering. These effects, particularly on the mitochondrial and plasma membranes, have been proposed to trigger toxicity of bile acids. This chapter details different fluorescence-based methods (fluorescence polarization, and spectroscopy/imaging of solvatochromic dyes) to evaluate the impact of different bile acids on membrane order. Protocols are described for the application of these methods to biomimetic vesicles, isolated mitochondria, and hepatocytes, following a bottom-up approach.

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Author information

Authors and Affiliations

  1. Centro de Química-Física Molecular and Institute of Nanoscience and Nanotechnology, Instituto Superior Técnico, University of Lisbon, Lisbon, Portugal

    Tânia Sousa, Ana Coutinho, Manuel Prieto & Fábio Fernandes

  2. iBB-Institute for Bioengineering and Biosciences, Biological Sciences Research Group, Lisbon, Portugal

    Tânia Sousa, Ana Coutinho, Manuel Prieto & Fábio Fernandes

  3. Faculty of Pharmacy, Research Institute for Medicines (iMed.ULisboa), University of Lisbon, Lisbon, Portugal

    Rui E. Castro & Cecília M. P. Rodrigues

  4. Faculty of Sciences, Departamento de Química e Bioquímica, University of Lisbon, Lisbon, Portugal

    Ana Coutinho

Authors
  1. Tânia Sousa
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  2. Rui E. Castro
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  3. Ana Coutinho
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  4. Cecília M. P. Rodrigues
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  5. Manuel Prieto
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  6. Fábio Fernandes
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Corresponding author

Correspondence to Fábio Fernandes .

Editor information

Editors and Affiliations

  1. Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Brussels, Belgium

    Mathieu Vinken

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Sousa, T., Castro, R.E., Coutinho, A., Rodrigues, C.M.P., Prieto, M., Fernandes, F. (2019). Measuring the Impact of Bile Acids on the Membrane Order of Primary Hepatocytes and Isolated Mitochondria by Fluorescence Imaging and Spectroscopy. In: Vinken, M. (eds) Experimental Cholestasis Research. Methods in Molecular Biology, vol 1981. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9420-5_7

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  • DOI: https://doi.org/10.1007/978-1-4939-9420-5_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9419-9

  • Online ISBN: 978-1-4939-9420-5

  • eBook Packages: Springer Protocols

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