TY - JOUR
T1 - Isolating Synaptic Vesicles from Neurospheres for Proteomics
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
UR - http://www.scopus.com/inward/record.url?scp=85213914665&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-4298-6_14
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 -