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Membrane Trafficking

Methods and Protocols

  • Book
  • © 2022

Overview

Editors:
  1. Jingshi Shen

    1. Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, USA

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

Part of the book series: Methods in Molecular Biology (MIMB, volume 2473)

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About this book

This detailed volume presents a series of methods exploring membrane trafficking research, ranging from genetics and high-resolution imaging to in vitro biochemical and biophysical assays. Covering virtually all the major trafficking branches, the book delves into the exocytic pathway, which focuses on cargo transport from the ER to the Golgi, through the Golgi cisternae, and to the plasma membrane and the extracellular space; the endocytic pathway, which includes cargo endocytosis, endosomal recycling, and lysosomal degradation; as well as emerging topics beyond the conventional exocytic and endocytic pathways. Written for the highly successful Methods in Molecular Biology series, 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 practical, Membrane Trafficking: Methods and Protocols provides techniques with broad applications as an ideal guide for junior researchers new to membrane trafficking as well as established membrane biologists seeking to expand their research programs.

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Keywords

Table of contents (25 protocols)

  1. Front Matter

    Pages i-xiv
    Download chapter PDF

  2. The Exocytic Pathway

    1. Front Matter

      Pages 1-1
      Download chapter PDF

    2. Characterizing Membrane Traffic in the Early Secretory Pathway Using the RUSH Retention System

      • Marine D. Camus, Stephane M. Camus
      Pages 3-14

    3. Monitoring Intra-Golgi Transport with Acute Spatiotemporal Control of a Synthetic Cargo

      • Julia Dancourt, Grégory Lavieu
      Pages 15-22

    4. Imaging Secretory Granule Budding from the Trans-Golgi Network Using Retention Using Selective Hook (RUSH)

      • Cedric S. Asensio
      Pages 23-28

    5. Spatial and Temporal Control of Protein Secretion with Light

      • Ashley M. Bourke, Matthew J. Kennedy
      Pages 29-45

    6. Generation of Endogenously Tagged Membrane Trafficking Regulators Using CRISPR Genome Editing

      • Syed Mukhtar Ahmed
      Pages 47-63

    7. Determine the Function of the Exocyst in Vesicle Tethering by Ectopic Targeting

      • Kunrong Mei, Di-Ao Liu, Wei Guo
      Pages 65-77

    8. Imaging Insulin Granule Dynamics in Human Pancreatic β-Cells Using Total Internal Reflection Fluorescence (TIRF) Microscopy

      • Fei Kang, Herbert Y. Gaisano
      Pages 79-88

    9. Monitoring Synaptic Exocytosis Using a Whole-Cell Patch Clamp Technique

      • Jihong Gong, Xiaofei Yang, Cong Ma
      Pages 89-100

    10. Live-Cell Superresolution Imaging of Retrograde Axonal Trafficking Using Pulse–Chase Labeling in Cultured Hippocampal Neurons

      • Tong Wang, Frédéric A. Meunier
      Pages 101-128

    11. Electron Tomographic Methods for Studying Organelles of the Murine Chemical Synapse

      • Tamara Basta, Garry P. Morgan, Eileen T. O’Toole, Nalini R. Rao, Jeffrey N. Savas, Michael H. B. Stowell
      Pages 129-139

    12. In Vitro Reconstitution Studies of SNAREs and Their Regulators Mediating GLUT4 Vesicle Fusion

      • Yinghui Liu, Ruyue He, Min Zhu, Haijia Yu
      Pages 141-156

    13. Imaging Single-Vesicle Exocytosis with Total Internal Reflection Fluorescence Microscopy (TIRFM)

      • Yingke Xu, Luhong Jin, Derek Toomre
      Pages 157-164

  3. The Endocytic Pathway

    1. Front Matter

      Pages 165-165
      Download chapter PDF

    2. Localizing Proteins on Single Trafficking Organelles in 3D with Semisynthetic Gold Labeling and Platinum Replica Electron Microscopy

      • Bijeta Prasai, Justin W. Taraska
      Pages 167-180

    3. Endocytosis Assays Using Cleavable Fluorescent Dyes

      • Shifeng Wang, Chun Wan, Galen T. Squiers, Jingshi Shen
      Pages 181-194

    4. Reconstituting and Purifying Assembly Intermediates of Clathrin Adaptors AP1 and AP2

      • Bing Wang, Rui Yang, Yuan Tian, Qian Yin
      Pages 195-212

    5. Monitoring Endosomal Cargo Retrieval to the Trans-Golgi Network by Microscopic and Biochemical Approaches

      • Amulya Priya, Sunando Datta
      Pages 213-236

    6. Measuring Plasma Membrane Recycling Using Microscopic and Biochemical Approaches

      • Priyanka Sharma, Sameena Parveen, Sunando Datta
      Pages 237-257
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Editors and Affiliations

  • Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, USA

    Jingshi Shen

Bibliographic Information

  • Book Title: Membrane Trafficking

  • Book Subtitle: Methods and Protocols

  • Editors: Jingshi Shen

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-0716-2209-4

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2022

  • Hardcover ISBN: 978-1-0716-2208-7Published: 13 July 2022

  • Softcover ISBN: 978-1-0716-2211-7Published: 13 July 2023

  • eBook ISBN: 978-1-0716-2209-4Published: 12 July 2022

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XIV, 407

  • Number of Illustrations: 9 b/w illustrations, 82 illustrations in colour

  • Topics: Membrane Biology, Cell Biology

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