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Series : Methods in Molecular Biology

Exocytosis and Endocytosis, Vol. 440, Ed. 1

¥ 8,393 (tax included)

Description

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.

Contents

Part I An Overview of Methods for Analyzing Vesicular Trafficking

1 Vesicular Trafficking: Molecular Tools and Targets

Elena Vassilieva and Asma Nusrat

2 Pharmacological Inhibition of Endocytic Pathways. Is It Specific Enough to be Useful?

Andrei Ivanov


Part II Cell-free and Biochemical Assays of Exocytosis and Endocytosis

3 In Vitro Assays to Measure SNARE-Mediated Vesicle Fusion

Susanne Kreye, Jorg Malsam, and Thomas H. Sollner

4 Imaging Ca2+-Triggered Exocytosis of Single Secretory Granules on Plasma Membrane Lawns from Neuroendocrine Cells.

Thorsten Lang

5 FRAP Analysis of Secretory Granule Lipids and Proteins in the Sea Urchin Egg

Julian L. Wong and Gary M. Wessel

6 A Cell-Free Assay for Endocytosis of E-cadherin

Toshiaki Sakisaka and Yoshimi Takai

7 Cell-Surface Biotinylation to Study Endocytosis and Recycling of Occludin

Noriyuki Nishimura and Takuya Sasaki

8 Fractionation of Subcellular Membrane Vesicles of Epithelial and Non-epithelial Cells by OptiPrepTM Density Gradient Ultracentrifugation

Xuhang Li and Mark Donowitz

9 Identification and Characterization of Interacting Partners of Rab GTPases by Yeast Two-Hybrid Analyses

Mark Kail and Angelika Barnekow


Part III Analysis of Endocytosis and Exocytosis in Different Cell Systems

10 High-Throughput Analysis of the Dynamics of Recycling Cell Surface Proteins

Roland Govers, David E. James, and Adelle C. F. Coster

11 Studying Phagocytosis by Live-Cell Scintillation Proximity Assay

Walter Stockinger and Axel Nohturfft

12 Transcytosis of Polymeric Immunoglobulin A in Polarized Madin-Darby Canine Kidney Cells

Asli Oztan, Christine Rondanino, and Gerard Apodaca

13 The Use of Syntaxin Chimeras to Study Polarized Protein Trafficking in Epithelial Cells

Martin B.A. ter Beest

14 Targeting the Epithelial SNARE Machinery by Bacterial Neurotoxins

Veronique Proux-Gillardeaux and Thierry Galli

15 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, and Charles J. Lowenstein

16 Molecular Dissection of HCl Secretion in Gastric Parietal Cells Using Streptolysin O Permeabilization

Xia Ding, Fang Wu, Zhen Guo, and Xuebiao Yao

17 Addressing Membrane Protein Topology Using the Fluorescence Protease Protection (FPP) Assay

Holger Lorenz, Dale W. Hailey, and Jennifer Lippincott-Schwartz

18 Visualizing Clathrin-Mediated IgE Receptor Internalization by Electron and Atomic Force Microscopy

Alan R. Burns, Janet M. Oliver, Janet R. Pfeiffer, and Bridget S. Wilson

19 Analyzing Endosomes in Non-Sectioned Cells by Transmission Electron Microscopy Willem Stoorvogel

20 Imaging Exocytosis of Single Insulin Secretory Granules with TIRF Microscopy

Shinya Nagamatsu and Mica Ohara-Imaizumi

21 Reconstitution of Depolarization and Ca2+-Evoked Secretion in Xenopus Oocytes Monitored by Membrane Capacitance

Roy Cohen, Bernhard M. Schmitt, and Daphne Atlas

22 Low-Noise Recording of Single Vesicle Capacitance Steps in Cell-Attached Patches

Vitaly Klyachko, Zhenjie Zhang, and Meyer Jackson

23 Good Practices in Single Cell Amperometry

J. David Machado, Monica S. Montesinos, and Ricardo Borges

24 Analysis of Single-Vesicle Exocytotic Events Recorded by Amperometry

Eugene V. Mosharov


Part IV. Analysis of Exocytosis and Endocytosis in Model Organisms

25 Application of RNAi Technology and Fluorescent Protein Markers to Study Membrane Traffic in C. elegans

Dmitry Poteryaev and Anne Spang

26 FM 1-43 Labeling of Synaptic Vesicle Pools at the Drosophila Neuromuscular Junction

Patrik Verstreken, Tokomo Ohyama, and Hugo J. Bellen

27 Probing E-cadherin Endocytosis by Morpholino-Mediated Rab5 Knock-Down in Zebrafish

Florian Ulrich and Carl-Philipp Heisenberg

28 Quantifying Endocytosis in vivo Using Intravital Two-Photon Microscopy

Ruben M. Sandoval and Bruce A. Molitoris