Isolating Synaptic Vesicles from Neurospheres for Proteomics

Caroline Brandão-Teles; Giuliana S. Zuccoli; Marcelo Ganzella; Victor Corasolla Carregari; Linda Olsthoorn; Érica de Almeida Duque; Carolina Demarchi Munhoz; Reinhard Jahn; Daniel Martins-de-Souza; Fernanda Crunfli

doi:10.1007/978-1-0716-4298-6_14

Isolating Synaptic Vesicles from Neurospheres for Proteomics

Caroline Brandão-Teles^*, Giuliana S. Zuccoli, Marcelo Ganzella, Victor Corasolla Carregari, Linda Olsthoorn, Érica de Almeida Duque, Carolina Demarchi Munhoz, Reinhard Jahn, Daniel Martins-de-Souza, Fernanda Crunfli^*

^*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Communication

Abstract

This chapter presents an optimized method for isolating synaptic vesicles (SVs) from neurospheres derived from human induced pluripotent stem cells (hiPSCs). The protocol begins with neurosphere cultivation to achieve mature neurons, which is essential for the functional studies of neuronal activity. Following this, neurosphere-derived synaptosomes are isolated, and SVs are enriched through differential centrifugation. The method culminates in the proteomic analysis of SVs using nano-liquid chromatography coupled with high-resolution tandem mass spectrometry (nanoLC-MS/MS), providing a detailed proteome profile of the isolated vesicles. This protocol can contribute to the understanding of SV molecular heterogeneity and the mechanisms of neurotransmitter uptake and release and be applied to the field of research in neurological and neuropsychiatric disorders.

Original language	English
Journal	Methods in Molecular Biology
Pages (from-to)	207-223
Number of pages	17
ISSN	1064-3745
DOIs	https://doi.org/10.1007/978-1-0716-4298-6_14
Publication status	Published - 2025

Keywords

Neural stem cell
Neurospheres
Proteomics
Synapse vesicles
Vesicle isolation

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10.1007/978-1-0716-4298-6_14

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title = "Isolating Synaptic Vesicles from Neurospheres for Proteomics",

abstract = "This chapter presents an optimized method for isolating synaptic vesicles (SVs) from neurospheres derived from human induced pluripotent stem cells (hiPSCs). The protocol begins with neurosphere cultivation to achieve mature neurons, which is essential for the functional studies of neuronal activity. Following this, neurosphere-derived synaptosomes are isolated, and SVs are enriched through differential centrifugation. The method culminates in the proteomic analysis of SVs using nano-liquid chromatography coupled with high-resolution tandem mass spectrometry (nanoLC-MS/MS), providing a detailed proteome profile of the isolated vesicles. This protocol can contribute to the understanding of SV molecular heterogeneity and the mechanisms of neurotransmitter uptake and release and be applied to the field of research in neurological and neuropsychiatric disorders.",

keywords = "Neural stem cell, Neurospheres, Proteomics, Synapse vesicles, Vesicle isolation",

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note = "Publisher Copyright: {\textcopyright} The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2025.",

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doi = "10.1007/978-1-0716-4298-6_14",

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journal = "Methods in Molecular Biology",

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AU - Brandão-Teles, Caroline

AU - Zuccoli, Giuliana S.

AU - Ganzella, Marcelo

AU - Corasolla Carregari, Victor

AU - Olsthoorn, Linda

AU - de Almeida Duque, Érica

AU - Munhoz, Carolina Demarchi

AU - Jahn, Reinhard

AU - Martins-de-Souza, Daniel

AU - Crunfli, Fernanda

N1 - Publisher Copyright: © The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2025.

PY - 2025

Y1 - 2025

N2 - This chapter presents an optimized method for isolating synaptic vesicles (SVs) from neurospheres derived from human induced pluripotent stem cells (hiPSCs). The protocol begins with neurosphere cultivation to achieve mature neurons, which is essential for the functional studies of neuronal activity. Following this, neurosphere-derived synaptosomes are isolated, and SVs are enriched through differential centrifugation. The method culminates in the proteomic analysis of SVs using nano-liquid chromatography coupled with high-resolution tandem mass spectrometry (nanoLC-MS/MS), providing a detailed proteome profile of the isolated vesicles. This protocol can contribute to the understanding of SV molecular heterogeneity and the mechanisms of neurotransmitter uptake and release and be applied to the field of research in neurological and neuropsychiatric disorders.

AB - This chapter presents an optimized method for isolating synaptic vesicles (SVs) from neurospheres derived from human induced pluripotent stem cells (hiPSCs). The protocol begins with neurosphere cultivation to achieve mature neurons, which is essential for the functional studies of neuronal activity. Following this, neurosphere-derived synaptosomes are isolated, and SVs are enriched through differential centrifugation. The method culminates in the proteomic analysis of SVs using nano-liquid chromatography coupled with high-resolution tandem mass spectrometry (nanoLC-MS/MS), providing a detailed proteome profile of the isolated vesicles. This protocol can contribute to the understanding of SV molecular heterogeneity and the mechanisms of neurotransmitter uptake and release and be applied to the field of research in neurological and neuropsychiatric disorders.

KW - Neural stem cell

KW - Neurospheres

KW - Proteomics

KW - Synapse vesicles

KW - Vesicle isolation

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DO - 10.1007/978-1-0716-4298-6_14

M3 - Journal article

C2 - 39716006

AN - SCOPUS:85213914665

SN - 1064-3745

SP - 207

EP - 223

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

ER -

Isolating Synaptic Vesicles from Neurospheres for Proteomics

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Keywords

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