Methods for Analysis of Golgi Complex Function

Methods for Analysis of Golgi Complex Function

1st Edition - November 21, 2013

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  • Editors: Franck Perez, David Stephens
  • Hardcover ISBN: 9780124171640
  • eBook ISBN: 9780124171831

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Description

This new volume of Methods in Cell Biology looks at methods for analyzing of golgi complex function. Chapters cover such topics as in vitro reconstitution systems, fluorescence-based analysis of trafficking in mammalian cells and high content screening. With cutting-edge material, this comprehensive collection is intended to guide researchers for years to come.

Key Features

  • Covers sections on model systems and functional studies, imaging-based approaches and emerging studies
  • Chapters are written by experts in the field
  • Cutting-edge material

Readership

Researchers and students in cell, molecular and developmental biology

Table of Contents

  • Series Page

    Contributors

    Preface

    Part 1: Model Systems and Functional Studies

    Chapter 1. Analysis of Golgi Complex Functions: In Vitro Reconstitution Systems

    Abstract

    Introduction

    Acknowledgments

    References

    Chapter 2. RNA Interference Approaches to Examine Golgi Function in Animal Cell Culture

    Abstract

    Overview of Gene Depletion Methodologies

    2.1 Key Considerations for the Analysis of Organelle Function

    2.2 Strategies for RNAi Experiments

    2.3 Choosing Between siRNA and shRNA

    2.4 Sources of Reagents for RNAi

    2.5 Controls

    2.6 Methods

    2.7 Discussion

    Summary

    Acknowledgments

    References

    Chapter 3. Trafficking Along the Secretory Pathway in Drosophila Cell Line and Tissues: A Light and Electron Microscopy Approach

    Abstract

    Introduction and Rationale

    3.1 Materials

    3.2 Methods

    Conclusions/Perspectives

    Acknowledgments

    References

    Chapter 4. Studying Membrane Trafficking in the Worm C. elegans by RNA Interference

    Abstract

    Introduction and Rationale

    4.1 Materials

    4.2 Methods

    4.3 Discussion

    Summary

    Acknowledgment

    References

    Chapter 5. Endomembrane and Golgi Traffic in Plant Cells

    Abstract

    Introduction

    5.1 Leaves as a Model System for Analyzing the Plant Golgi and Secretory Pathway

    5.2 Mutant Screens for Trafficking Proteins

    5.3 Photobleaching for Transport Studies

    5.4 Förster Resonance Energy Transfer for Protein Interactions

    Summary

    Acknowledgments

    References

    Chapter 6. Biochemical Analysis of Secretory Trafficking in Mammalian Cells

    Abstract

    Introduction

    6.1 Materials

    6.2 Methods

    6.3 Discussion

    Summary

    Acknowledgments

    References

    Chapter 7. Methods for Analysis of Apical Lumen Trafficking Using Micropatterned 3D Systems

    Abstract

    Introduction

    7.1 3D Culture of MDCK Cysts on Micropatterns

    7.2 Discussion

    Acknowledgments

    References

    Chapter 8. Using Replication Defective Viruses to Analyze Membrane Trafficking in Polarized Epithelial Cells

    Abstract

    Introduction and Rationale

    8.1 Materials

    8.2 Methods

    8.3 Discussion

    Summary

    Acknowledgments

    References

    Chapter 9. SNAP-Tagging the Retrograde Route

    Abstract

    Introduction

    9.1 Materials

    9.2 Methods

    9.3 Conclusion/Discussion

    Summary

    Acknowledgments

    References

    Chapter 10. Assessing ER and Golgi N-Glycosylation Process Using Metabolic Labeling in Mammalian Cultured Cells

    Abstract

    Introduction and Rationale

    10.1 Materials

    10.2 Methods

    10.3 Discussion

    Summary

    Acknowledgments

    References

    Part 2: Imaging-Based Approaches

    Chapter 11. Fluorescence-Based Analysis of Trafficking in Mammalian Cells

    Abstract

    Introduction

    11.1 Methods

    11.2 Discussion

    Summary

    Acknowledgments

    References

    Chapter 12. Photobleaching Methods to Study Golgi Complex Dynamics in Living Cells

    Abstract

    Introduction and Rationale

    Summary

    Acknowledgments

    References

    Chapter 13. Photohighlighting Approaches to Access Membrane Dynamics of the Golgi Apparatus

    Abstract

    Introduction

    Summary

    References

    Chapter 14. Live Cell Visualization of Golgi Membrane Dynamics by Super-resolution Confocal Live Imaging Microscopy

    Abstract

    Introduction

    Acknowledgments

    References

    Chapter 15. Analysis of Golgi Complex Function Using Correlative Light-Electron Microscopy

    Abstract

    Introduction

    15.1 Materials

    15.2 Methods

    15.3 Discussion

    Acknowledgments

    References

    Chapter 16. Viewing Golgi Structure and Function from a Different Perspective—Insights from Electron Tomography

    Abstract

    Abbreviations

    Introduction

    Summary

    Acknowledgments

    References

    Chapter 17. High-Content Screening and Analysis of the Golgi Complex

    Abstract

    Introduction

    17.1 Materials

    17.2 Methods

    17.3 Discussion

    Summary

    Acknowledgments

    References

    Part 3: Emerging Studies [New Frontiers]

    Chapter 18. (Re)Modeling the Golgi

    Abstract

    Introduction: Coarse-Grained Modeling Based on Nonequilibrium Physics

    Acknowledgments

    References

    Chapter 19. Golgi Depletion from Living Cells with Laser Nanosurgery

    Abstract

    Introduction and Rationale

    19.1 Theory

    19.2 Equipment

    19.3 Materials

    19.4 Methods

    19.5 Discussion

    Acknowledgment

    References

    Chapter 20. Studying Intracellular Trafficking Pathways with Probabilistic Density Maps

    Abstract

    Introduction

    20.1 Material and Methods

    20.2 Results and Discussion

    Summary

    Acknowledgments

    References

    Chapter 21. Methods to Study Signaling at the Golgi Apparatus

    Abstract

    Introduction and Rationale

    21.1 Materials

    21.2 Methods

    Concluding Remarks

    Acknowledgments

    References

    Chapter 22. Golgi-Dependent Signaling: Self-Coordination of Membrane Trafficking

    Abstract

    Abbreviations

    Introduction

    22.1 Generation of Traffic-Dependent Golgi Signaling: ER-to-Golgi Traffic Synchronization Protocols

    22.2 Morphological and Quantitative Analysis of Traffic-Generated Golgi-Based Signaling

    22.3 Analysis of KDELR-Dependent Signaling

    22.4 The KDELR Transduction Machinery

    22.5 Discussion

    Summary

    Acknowledgment

    References

    Chapter 23. Signaling at the Golgi During Mitosis

    Abstract

    Introduction

    23.1 The Organization of Mammalian Golgi Membranes in Interphase

    23.2 Mechanism of Mitotic Golgi Disassembly

    23.3 Mechanism of Postmitotic Golgi Reassembly

    23.4 The Significance of Golgi Reorganization in Mitosis

    23.5 Assays to Study Mitotic Golgi Reorganization

    Concluding Remarks

    Acknowledgments

    References

    Chapter 24. Ice Recovery Assay for Detection of Golgi-Derived Microtubules

    Abstract

    Introduction and Rationale

    Summary

    Acknowledgments

    References

    Index

    Volumes in Series

Product details

  • No. of pages: 498
  • Language: English
  • Copyright: © Academic Press 2013
  • Published: November 21, 2013
  • Imprint: Academic Press
  • Hardcover ISBN: 9780124171640
  • eBook ISBN: 9780124171831

About the Serial Volume Editors

Franck Perez

Affiliations and Expertise

Institut Curie, CNRS, France

David Stephens

School of Biochemistry, Bristol University, UK

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