Exocytosis and Endocytosis

by Andrei I. Ivanov (Editor)

Overview

Due to their vital involvement in a wide variety of housekeeping and specialized cellular functions, exocytosis and endocytosis remain among the most popular subjects in biology and biomedical sciences. Tremendous progress in understanding these complex intracellular processes has been achieved by employing a wide array of research tools ranging from classical biochemical methods to modern imaging techniques. In Exocytosis and Endocytosis, skilled experts provide the most up-to-date, step-by-step laboratory protocols for examining molecular machinery and biological functions of exocytosis and endocytosis in vitro and in vivo. Following the highly successful Methods in Molecular Biology series format, the chapters present an introduction outlining the principle behind each technique, a list of the necessary materials, an easy to follow, readily reproducible protocol, and a Notes section offering tips on troubleshooting and avoiding known pitfalls.

Insightful to both newcomers and seasoned professionals, Exocytosis and Endocytosis offers a unique and highly practical guide to versatile laboratory tools developed to study various aspects of intracellular vesicle trafficking in simple model systems and living organisms.

Product Details

ISBN-13:	9781493909438
Publisher:	Springer New York
Publication date:	06/20/2014
Series:	Methods in Molecular Biology , #1174
Edition description:	2nd ed. 2014
Pages:	435
Product dimensions:	7.01(w) x 10.00(h) x 0.04(d)

Table of Contents

Preface     v
Contributors     xi
An Overview of Methods for Analyzing Vesicular Trafficking
Vesicular Trafficking: Molecular Tools and Targets   Elena V. Vassilieva   Asma Nusrat     3
Pharmacological Inhibition of Endocytic Pathways: Is It Specific Enough to Be Useful?   Andrei I. Ivanov     15
Cell-Free and Biochemical Assays of Exocytosis and Endocytosis
In Vitro Assays to Measure SNARE-Mediated Vesicle Fusion   Susanne Kreye   Jorg Malsam   Thomas H. Sollner     37
Imaging Ca[superscript 2+]-Triggered Exocytosis of Single Secretory Granules on Plasma Membrane Lawns from Neuroendocrine Cells   Thorsten Lang     51
FRAP Analysis of Secretory Granule Lipids and Proteins in the Sea Urchin Egg   Julian L. Wong   Gary M. Wessel     61
A Cell-Free Assay for Endocytosis of E-Cadherin   Toshiaki Sakisaka   Yoshimi Takai     77
Cell-Surface Biotinylation to Study Endocytosis and Recycling of Occludin   Noriyuki Nishimura   Takuya Sasaki     89
Fractionation of Subcellular Membrane Vesicles of Epithelial and Nonepithelial Cells by OptiPrep Density Gradient Ultracentrifugation   Xuhang Li   Mark Donowitz     97
Identification and Characterization of Interacting Partners of Rab GTPases by YeastTwo-Hybrid Analyses   Mark Kail   Angelika Barnekow     111
Analysis of Endocytosis and Exocytosis in Different Cell Systems
High-Throughput Analysis of the Dynamics of Recycling Cell Surface Proteins   Roland Govers   David E. James   Adelle C.F. Coster     129
Studying Phagocytosis by Live-Cell Scintillation Proximity Assay   Walter Stockinger   Axel Nohturfft     147
Transcytosis of Polymeric Immunoglobulin A in Polarized Madin-Darby Canine Kidney Cells   Asli Oztan   Christine Rondanino   Gerard Apodaca     157
The Use of Syntaxin Chimeras to Study Polarized Protein Trafficking in Epithelial Cells   Martin B.A. ter Beest     171
Targeting the Epithelial SNARE Machinery by Bacterial Neurotoxins   Veronique Proux-Gillardeaux   Thierry Galli     187
Exocytosis of Endothelial Cells Is Regulated by N-Ethylmaleimide-Sensitive Factor   Munekazu Yamakuchi   Marcella Ferlito   Craig N. Morrell   Kenji Matsushita   Craig A. Fletcher   Wangsen Cao   Charles J. Lowenstein     203
Molecular Dissection of HCl Secretion in Gastric Parietal Cells Using Streptolysin O Permeabilization   Xia Ding   Fang Wu   Zhen Guo   Xuebiao Yao      217
Addressing Membrane Protein Topology Using the Fluorescence Protease Protection (FPP) Assay   Holger Lorenz   Dale W. Hailey   Jennifer Lippincott-Schwartz     227
Visualizing Clathrin-Mediated IgE Receptor Internalization by Electron and Atomic Force Microscopy   Alan R. Burns   Janet M. Oliver   Janet R. Pfeiffer   Bridget S. Wilson     235
Analyzing Endosomes in Nonsectioned Cells by Transmission Electron Microscopy   Willem Stoorvogel     247
Imaging Exocytosis of Single Insulin Secretory Granules With TIRF Microscopy   Shinya Nagamatsu   Mica Ohara-Imaizumi     259
Reconstitution of Depolarization and Ca[superscript 2+]-Evoked Secretion in Xenopus Oocytes Monitored by Membrane Capacitance   Roy Cohen   Bernhard M. Schmitt   Daphne Atlas     269
Low-Noise Recording of Single-Vesicle Capacitance Steps in Cell-Attached Patches   Vitaly Klyachko   Zhenjie Zhang   Meyer Jackson     283
Good Practices in Single-Cell Amperometry   David J. Machado   Monica S. Montesinos   Ricardo Borges     297
Analysis of Single-Vesicle Exocytotic Events Recorded by Amperometry   Eugene V. Mosharov     315
Analysis of Exocytosis and Endocytosis in Model Organisms
Application of RNAi Technology and Fluorescent Protein Markers to Study Membrane Traffic in Caenorhabditis elegans   Dmitry Poteryaev   Anne Spang     331
FM 1-43 Labeling of Synaptic Vesicle Pools at the Drosophila Neuromuscular Junction   Patrik Verstreken   Tomoko Ohyama   Hugo J. Bellen     349
Probing E-Cadherin Endocytosis by Morpholino-Mediated Rab5 Knockdown in Zebrafish   Florian Ulrich   Carl-Philipp Heisenberg     371
Quantifying Endocytosis In Vivo Using Intravital Two-Photon Microscopy   Ruben M. Sandoval   Bruce A. Molitoris     389
Index     403