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Isolation of Synaptic Vesicles from Mammalian Brain

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Synaptic Vesicles

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2417))

Abstract

Synaptic vesicles (SVs) store neurotransmitters and undergo a fine-tuned regulatory and dynamic cycle of exo- and endocytosis, which is essential for neurotransmission at chemical synapses. The development of protocols for isolating SVs from biological extracts was a fundamental accomplishment since it allowed for characterizing the molecular properties of SVs using biochemical methods. In this chapter, we describe a modified procedure for isolating SVs from a few g of rodent brain and that can be completed within ~12 h. The protocol involves the preparation of isolated nerve terminals from which SVs are released by osmotic shock and then enriched via various centrifugation steps, followed by size exclusion chromatography as final purification step. The final vesicle fraction is 22-fold enriched in SVs over the starting material, and the final yield of SVs obtained using this protocol is approximately 20 μg of protein per gram of mouse brain. The degree of contamination by other organelles and particles monitored by morphology and immunolabeling compares well with that of the classical protocols.

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Acknowledgments

We would like to sincerely thank Miroslav Gaydarski for the support with the graphical formulation of the figures. We are also thankful to Mehar Monga and Gudrun Heim for the excellent technical assistance.

Author information

Authors and Affiliations

  1. Laboratory of Neurobiology, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany

    Marcelo Ganzella, Momchil Ninov & Reinhard Jahn

  2. Bioanalytical Mass Spectrometry, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany

    Momchil Ninov

  3. Facility for Transmission Electron Microscopy, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany

    Dietmar Riedel

  4. Georg-August University, Göttingen, Germany

    Reinhard Jahn

Authors
  1. Marcelo Ganzella
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  2. Momchil Ninov
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  3. Dietmar Riedel
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  4. Reinhard Jahn
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Corresponding author

Correspondence to Marcelo Ganzella .

Editor information

Editors and Affiliations

  1. Department of Psychiatry and Psychotherapy, Universitätsklinikum Erlangen, Erlangen, Bayern, Germany

    Jana Dahlmanns

  2. Institute for Physiology and Pathophysiology, University of Erlangen-Nuremberg, Erlangen, Germany

    Marc Dahlmanns

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Ganzella, M., Ninov, M., Riedel, D., Jahn, R. (2022). Isolation of Synaptic Vesicles from Mammalian Brain. In: Dahlmanns, J., Dahlmanns, M. (eds) Synaptic Vesicles. Methods in Molecular Biology, vol 2417. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1916-2_11

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  • DOI: https://doi.org/10.1007/978-1-0716-1916-2_11

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1915-5

  • Online ISBN: 978-1-0716-1916-2

  • eBook Packages: Springer Protocols

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