Main description:

This book provides protocols on the preparation of synaptic vesicles from different model systems and provides instructions for functional investigations of their transport and formation, scaffolding, filling and priming, release, recycling, and degradation. Chapters guide readers through computational approaches, simulating different aspects of synaptic vesicle anatomy and function. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

Authoritative and cutting-edge, Synaptic Vesicles: Methods and Protocols aims to be a useful practical guide to researchers to help further their study in this field.

Contents:

1. 1. Real-time quantitative PCR and fluorescence in situ hybridization for subcellular localization of miRNAs in neurons.

Antonis Tatarakis and Danesh Moazed

2. Live-imaging of axonal cargoes in Drosophila brain explants using confocal microscopy.
Caroline Medioni, Anne Ephrussi and Florence Besse

3. Whole Endosome Recording of Vesicular Neurotransmitter Transporter Currents.
Roger Chang and Robert H. Edwards

4. Quantitative analysis of presynaptic vesicle luminal pH in cultured neurons.
Yoshihiro Egashira, Shutaro Katsurabayashi and Shigeo Takamori

5. Investigation of synaptic vesicle proteins in rat brain tissue using real-time qPCR.
Betina Elfving

6. Mass Synaptometry: Applying Mass Cytometry to Single Synapse Analysis.
Chandresh R. Gajera, Rosemary Fernandez, Nadia Postupna, Kathleen S. Montine, C. Dirk Keene, Sean C. Bendall, Thomas J. Montine

7. A Guide to Analysis of Relative Synaptic Protein Abundance by Quantitative Fluorescent Western Blotting.
Heidi Kaastrup Muller

8. Synaptosomes and metamodulation of receptors.
Anna Pittaluga and Mario Marchi

9. A novel method to monitor GABA loading into synaptic vesicles by combining patch pipette perfusion and intracellular, caged-GABA photolysis in brain slice preparations.
Manami Yamashita and Tetsuya Hori

10. Rapid Isolation of Functional Synaptic Vesicles from Tissues through Cryo-grinding, Ultracentrifugation and Size Exclusion Chromatography.
Huinan Marcus Li

11. Isolation of synaptic vesicles from mammalian brain
Marcelo Ganzella, Momchil Ninov, Dietmar Riedel, Reinhard Jahn

12. Building and Using a Two-Photon Fluorescence Crosscorrelation Spectroscopy Setup including Fluorescence Lifetime Analysis.
Tobias Grothe and Peter J. Walla

13. Fluorescence Lifetime and Cross-correlation Spectroscopy for Observing Membrane Fusion of Liposome Models Containing Synaptic Proteins.
Tobias Grothe and Peter J. Walla

14. Synaptic vesicle pool monitoring with synaptopH-Iuorin.
Marc Dahlmanns and Jana Katharina Dahlmanns

15. Imaging neuropeptide release at Drosophila neuromuscular junction with a genetically engineered neuropeptide release reporter.
Yifu Han and Keke Ding

16. Imaging synaptic glutamate release with two-photon microscopy in organotypic slice cultures
Celine D. Durst and Thomas G. Oertner

17. Dynole 34-2 and Acrylo-Dyn 2-30, novel dynamin GTPase chemical biology probes.
Jennifer R. Baker, Nicholas S. O'Brien, Kate L. Prichard, Phillip J. Robinson, Adam McCluskey and Cecilia C. Russell

18. Synthesis of Pthaladyn-29 and Napthalimide-10, GTP site directed dynamin GTPase inhibitors.
Cecilia C. Russell, Kate L. Prichard, Nicholas S. O'Brien, Adam McCluskey, Phillip J. Robinson and Jennifer R. Baker