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Caveolae

Methods and Protocols

  • Book
  • © 2020

Overview

Editors:
  1. Cedric M. Blouin

    1. Institut Curie – Centre de Recherche, PSL Research University, Membrane Mechanics and Dynamics of Intracellular Signaling Laboratory, Paris, France

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  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 2169)

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About this book

This volume explores techniques used to study either the structure or the functions of caveolae and their components in several normal and pathophysiological situations. The chapters in this book cover topics such as selective visualization of caveolae by electron microscopy techniques; spatiotemporal analysis of caveolae dynamics and mechanics using live cell fluorescence microscopy; in vitro reconstitution such as liposomes and GPMVs as tools to study caveolin- and cavin-interacting partners; pulling of tethers from the cell plasma membrane using optical tweezers; and immunofluorescence-based analysis of caveolin-3 in the diagnostic management of neuromuscular diseases. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and comprehensive, Caveolae: Methods and Protocols is a valuable resource for both novice and expert researchers who are interested in discovering new roles or regulations of formed caveolae, and proteins composing their coat in various model organisms.  


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Table of contents (18 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. Selective Visualization of Caveolae by TEM Using APEX2

    • Alexander Ludwig
    Pages 1-10

  3. Freeze-Fracture Replica Immunolabeling of Cryopreserved Membrane Compartments, Cultured Cells and Tissues

    • Eric Seemann, Michael M. Kessels, Britta Qualmann
    Pages 11-25

  4. Analysis of Caveolin in Primary Cilia

    • Stine K. Morthorst, Johanne B. Mogensen, Søren T. Christensen, Lotte B. Pedersen
    Pages 27-41

  5. Method for Efficient Observation of Caveolin-1 in Plasma Membrane by Microscopy Imaging Analysis

    • Tomoya Yamaguchi, Toyoshi Fujimoto, Takashi Takahashi
    Pages 43-52

  6. Quantitative Image Analysis of the Spatial Organization and Mobility of Caveolin Aggregates at the Plasma Membrane

    • Takashi Hirama, Raibatak Das
    Pages 53-62

  7. Spatiotemporal Analysis of Caveolae Dynamics Using Total Internal Reflection Fluorescence Microscopy

    • Yosuke Senju, Shiro Suetsugu
    Pages 63-70

  8. Live-Cell FRET Imaging of Phosphorylation-Dependent Caveolin-1 Switch

    • Adriana M. Zimnicka, Zhenlong Chen, Peter T. Toth, Richard D. Minshall
    Pages 71-80

  9. GPMVs as a Tool to Study Caveolin-Interacting Partners

    • Joanna Podkalicka, Cedric M. Blouin
    Pages 81-88

  10. Biotin Proximity Labeling to Identify Protein–Protein Interactions for Cavin1

    • Carolina Mendoza-Topaz
    Pages 89-103

  11. Investigation of Novel Cavin-1/Suppressor of Cytokine Signaling 3 (SOCS3) Interactions by Coimmunoprecipitation, Peptide Pull-Down, and Peptide Array Overlay Approaches

    • Jamie J. L. Williams, George S. Baillie, Timothy M. Palmer
    Pages 105-118

  12. Analysis of Protein and Lipid Interactions Using Liposome Co-sedimentation Assays

    • Elin Larsson, Madlen Hubert, Richard Lundmark
    Pages 119-127

  13. Liposome Binding Assay to Characterize the Structure and Function of Cavin Proteins

    • Miriam Stoeber
    Pages 129-136

  14. Preparation of Caveolin-1 for NMR Spectroscopy Experiments

    • Sarah M. Plucinsky, Jeffrey A. Julien, Kerney Jebrell Glover
    Pages 137-147

  15. Tagging and Deleting of Endogenous Caveolar Components Using CRISPR/Cas9 Technology

    • Elena Shvets, Carolina Mendoza-Topaz
    Pages 149-166

  16. Pulling of Tethers from the Cell Plasma Membrane Using Optical Tweezers

    • Darius V. Köster
    Pages 167-174

  17. Live Confocal Imaging of Zebrafish Notochord Cells Under Mechanical Stress In Vivo

    • Ye-Wheen Lim, Harriet P. Lo, Thomas E. Hall, Robert G. Parton
    Pages 175-187

  18. Study of Caveolae-Dependent Mechanoprotection in Human Muscle Cells Using Micropatterning and Live-Cell Microscopy

    • Melissa Dewulf, Cedric M. Blouin
    Pages 189-196

  19. Immunofluorescence-Based Analysis of Caveolin-3 in the Diagnostic Management of Neuromuscular Diseases

    • Andreas Roos, Denisa Hathazi, Ulrike Schara
    Pages 197-216

  20. Back Matter

    Pages 217-218
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Editors and Affiliations

  • Institut Curie – Centre de Recherche, PSL Research University, Membrane Mechanics and Dynamics of Intracellular Signaling Laboratory, Paris, France

    Cedric M. Blouin

Bibliographic Information

  • Book Title: Caveolae

  • Book Subtitle: Methods and Protocols

  • Editors: Cedric M. Blouin

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-0716-0732-9

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2020

  • Hardcover ISBN: 978-1-0716-0731-2Published: 17 June 2020

  • Softcover ISBN: 978-1-0716-0734-3Published: 17 June 2021

  • eBook ISBN: 978-1-0716-0732-9Published: 16 June 2020

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 218

  • Number of Illustrations: 9 b/w illustrations, 37 illustrations in colour

  • Topics: Lipidology, Cell Biology

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