Methods for Analysis of Golgi Complex Function

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