Membrane Trafficking, 1st ed. 2022 Methods and Protocols Methods in Molecular Biology Series, Vol. 2473

Part I: The Exocytic Pathway

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

            Marine D. Camus and Stephane M. Camus

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

            Julia Dancourt and Grégory Lavieu

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

            Cedric S. Asensio

4. Spatial and Temporal Control of Protein Secretion with Light

            Ashley M. Bourke and Matthew J. Kennedy

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

            Syed Mukhtar Ahmed

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

            Kunrong Mei, Di-Ao Liu, and Wei Guo

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

            Fei Kang and Herbert Y. Gaisano

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

            Jihong Gong, Xiaofei Yang, and Cong Ma

9. Live-Cell Super-Resolution Imaging of Retrograde Axonal Trafficking Using Pulse-Chase Labeling in Cultured Hippocampal Neurons

            Tong Wang and Frédéric A. Meunier

10. 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, and Michael H.B. Stowell

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

            Yinghui Liu, Ruyue He, Min Zhu, and Haijia Yu

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

Part II: The Endocytic Pathway

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

            Bijeta Prasai and Justin W. Taraska

14. Endocytosis Assays Using Cleavable Fluorescent Dyes

            Shifeng Wang, Chun Wan, Galen T. Squiers, and Jingshi Shen

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

            Bing Wang, Rui Yang, Yuan Tian, and Qian Yin

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

            Amulya Priya and Sunando Datta

17. Measuring Plasma Membrane Recycling Using Microscopic and Biochemical Approaches

            Priyanka Sharma, Sameena Parveen, and Sunando Datta

18. Light Scattering Techniques to Assess Self-Assembly and Hydrodynamics of Membrane Trafficking Proteins

            Marijn G.J. Ford and Rajesh Ramachandran

19. Measurement of Lysosome Positioning by Shell Analysis and Line Scan

            Chad D. Williamson, Carlos M. Guardia, Raffaella De Pace, Juan S. Bonifacino, and Amra Saric

Part III: Emerging Topics in Membrane Trafficking

20. Optogenetic Control of Membrane Trafficking Using Light-Activated Reversible Inhibition by Assembly Trap of Intracellular Membranes (IM-LARIAT)

            Bobae Hyeon, Mai Khanh Nguyen, and Won Do Heo

21. Generating Custom Pooled CRISPR Libraries for Genetic Dissection of Biological Pathways

22. Studying Unconventional Secretion of Misfolded Proteins in Cultured Cells and Primary Neurons

            Juhyung Lee and Yihong Ye

23. Single-Molecule Optical Tweezers Study of Protein-Membrane Interactions

24. Generation of Hybrid Extracellular Vesicles by Fusion with Functionalized Liposomes

            Max Piffoux, Amanda Karine Andriola Silva, Florence Gazeau, and David Tareste

25. Quantitative Measurement of Extracellular Vesicle Content Delivery within Acceptor Cells

            Émeline Bonsergent, Shéryl Bui, and Grégory Lavieu

Includes cutting-edge techniques

Provides step-by-step detail essential for reproducible results

Contains key implementation advice from the experts