Exocytosis and Endocytosis, 1st ed. 2021 Methods and Protocols Methods in Molecular Biology Series, Vol. 2233

This volume provides basic and cutting-edge methods and protocols to study the major characteristics of eukaryotic cells.  Chapters detail the different pathways of endocytosis in vivo, real time imaging of endocytic steps, endocytosis in model organisms, super-resolution methods to follow proteins involved in exocytosis, specific protocols for exocytosis in specialized cells such as neutrophils or neuroendocrine cells, as well as secretion of exosomes. 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.

Cutting-edge and clearly written, Exocytosis and Endocytosis: Methods and Protocols is a valuable resource for researchers in the fields of cell biology, neurology, immunology, oncology, and those interested in studying protein trafficking and signal regulation.

Part I. Endocytosis

1. Stoichiometry of Receptors at the Plasma Membrane during Their Endocytosis using Total Internal Reflection Fluorescent (TIRF) Microscopy Live Imaging and Single Molecule Tracking

Laura SalavessaandNathalie Sauvonnet

Claudia Hinze, Kieran McGourty, and Emmanuel Boucrot

3. Measurements of Compensatory Endocytosis by Antibody Internalization and Quantification of Endocytic Vesicle Distribution In Adrenal Chromaffin Cells

Mara Ceridono, Sylvette Chasserot-Golaz, Nicolas Vitale, Stéphane Gasman, and Stéphane Ory

4. Quantitative Methods to Study Endocytosis and Retrograde Transport of Cargo Proteins

David A. Cardoso, Ngoc Chau, and Phillip J. Robinson

6. Methods for Monitoring Endocytosis in Astrocytes

Maja Potokar, Jernej Jorgačevski, and Robert Zorec

7. Monitoring Activity-Dependent Bulk Endocytosis in Primary Neuronal Culture Using Large Fluorescent Dextrans

Michael A. Cousinand Karen J. Smillie

PART II. Exocytosis

8. Quantitative Flow Cytometry-Based Assays for Measuring Constitutive Secretion

David E. Gordon, Amber S. Shun-Shion, Asral W. Asnawi, and Andrew A. Peden

9. High-throughput Screening for Insulin Secretion Modulators

Michael A. Kalwat

10. Different Approaches to Record Human Sperm Exocytosis

Laila Suhaiman, Karina Noel Altamirano, Alfonsina Morales, and Silvia Alejandra Belmonte

11. Bovine Chromaffin Cells: Culture and Fluorescent Assay for Secretion

Thamou Thahouly, Emeline Tanguy, Juliette Raherindratsara, Marie-France Bader, Sylvette Chasserot-Golaz, Stéphane Gasman, and Nicolas Vitale

12. Measurement of Exocytosis in Genetically Manipulated Mast Cells
Ofir Klein, Nurit P Azouz, and Ronit Sagi-Eisenberg

13. Super-Resolution Microscopy and Particle Tracking Approaches for the Study of Vesicular Trafficking in Primary Neutrophils

Jennifer L. Johnson, Kersi Pestonjamasp, William B. Kiosses, and Sergio D. Catz

14. An Approach to Monitor Exocytosis in White Adipocytes

Ali M. Komai, Man Mohan Shrestha, Saliha Musovic, and Charlotta S. Olofsson

15. Amperometry in Single Cells and Tissue

Damien J. Keating

16. Measurements of Exocytosis by Capacitance Recordings and Calcium Uncaging In Mouse Adrenal Chromaffin Cells

Sébastien Houy, Joana S. Martins, Ralf Mohrmann, and Jakob Balslev Sørensen

17. RUSH-synchronized Secretion to Measure Local Exocytosis

Gaelle Boncompain, Lou Fourriere, Nelly Gareil, and Franck Perez

18. Combining Single Molecule Super-Resolution Imaging Techniques to Unravel the Nanoscale Organization of the Presynapse

Christopher Small, Ramon Martínez-Mármol, Rumelo Amor, Merja Joensuu, and Frederic A. Meunier

19. Induction of Ca²⁺ Dependent Exocytotic Processes by Laser Ablation of Endothelial Cells

Arsila P. K. Ashraf, Sophia N. Koerdt, Nikita Raj, and Volker Gerke

21. Spatial and Temporal Aspects 0f Exocytosis Studied on the Isolated Plasma Membranes

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts