Main description:

This book presents a methodological introduction on clathrin-mediated endocytosis. Chapters guide readers through a number of model systems, purifications, reconstitutions, analysis techniques and systemic alterations of membrane function, which has been pivotal in understanding the role of clathrin and is panoply of interactors. 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.

Authoritative and cutting-edge, Clathrin-Mediated Endoytsis: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Contents:

1. Purification of Clathrin-coated Vesicles from Adult Rat Brain

Silvia Giovedi

2. Preparation of Synaptosomes from Mammalian Brain by Subcellular Fractionation and Gradient Centrifugation

Mirko Messa

3. Probing Endocytosis during the Cell Cycle with Minimal Experimental Perturbation

Antonio JM Santos and Emmanuel Boucrot

4. Assaying the Contribution of Membrane Tension to Clathrin-mediated Endocytosis

Steeve Boulant

5Identifying Small Molecule Inhibitors of the Clathrin Terminal Domain

Volker Haucke and Michael Krauss

6. Acute Manipulations of Clathrin-mediated Endocytosis at Presynaptic Nerve Terminals

Rylie B. Walsh, Ona E. Bloom, and Jennifer R. Morgan

7. Imaging "Hot-wired" Clathrin-mediated Endocytosis

Laura A. Wood and Stephen J. Royle

8. Real-time Endocytosis Measurements by Membrane Capacitance Recording at Central Nerve Terminals

Xuelin Lou

9. Assaying Mutants of Clathrin Mediated Endocytosis in the Fly Eye

Elsa Lauwers and Patrik Verstreken

10. Reconstitution of Clathrin Coat Disassembly for Fluorescence Microscopy and Single-molecule Analysis

Till Boecking , Srigokul Upadhyayula, Iris Rapoport, Benjamin R. Capraro, and Tomas Kirchhausen

11. Spatial and Temporal Aspects of Phosphoinositides in Endocytosis Studied in Isolated Plasma Membranes

Ira Milosevic

12. SMrT Assay for Real-time Visualization and Analysis of Clathrin Assembly Reactions

Devika Andhare, Sachin S. Holkar, and Thomas J. Pucadyil

13. Real-time Monitoring of Clathrin Assembly Kinetics in a Reconstituted System

Jeffery Yong, Yan Chen, and Min Wu

14. Stimulated Emission Depletion (STED) Imaging of Clathrin-mediated Endocytosis in Living Cells

Francesca Bottanelli and Lena Schroeder

15. Measuring Clathrin-coated Vesicle Formation with Single-molecule Resolution

Francois Aguet and Emanuele Cocucci

16. Cryo-electron Tomography of the Mammalian Synapse

Ruben Fernandez-Busnadiego

17. Quantitative Analysis of Clathrin-mediated Endocytosis in Yeast by Live Cell Fluorescence Microscopy

Eric B Lewellyn and Yansong Miao

18. Using FM Dyes to Monitor Clathrin-mediated Endocytosis in Primary Neuronal Culture

Michael A. Cousin, Sarah L. Gordon, and Karen J. Smillie