Surface charge manipulation and electrostatic immobilization of synaptosomes for super-resolution imaging: a study on tau compartmentalization

Abstract Synaptosomes are subcellular fractions prepared from brain tissues that are enriched in synaptic terminals, widely used for the study of neural transmission and synaptic dysfunction. Immunofluorescence imaging is increasingly applied to synaptosomes to investigate protein localization. Howe...

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Autores principales:	Ushashi Bhattacharya, Jia-Fong Jhou, Yi-Fong Zou, Gerald Abrigo, Shu-Wei Lin, Yun-Hsuan Chen, Fan-Ching Chien, Hwan-Ching Tai
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/dcce73005ca6481c969d0e8d56ddfc6b
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spelling	oai:doaj.org-article:dcce73005ca6481c969d0e8d56ddfc6b2021-12-02T15:14:37ZSurface charge manipulation and electrostatic immobilization of synaptosomes for super-resolution imaging: a study on tau compartmentalization10.1038/s41598-021-98142-12045-2322https://doaj.org/article/dcce73005ca6481c969d0e8d56ddfc6b2021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-98142-1https://doaj.org/toc/2045-2322Abstract Synaptosomes are subcellular fractions prepared from brain tissues that are enriched in synaptic terminals, widely used for the study of neural transmission and synaptic dysfunction. Immunofluorescence imaging is increasingly applied to synaptosomes to investigate protein localization. However, conventional methods for imaging synaptosomes over glass coverslips suffer from formaldehyde-induced aggregation. Here, we developed a facile strategy to capture and image synaptosomes without aggregation artefacts. First, ethylene glycol bis(succinimidyl succinate) (EGS) is chosen as the chemical fixative to replace formaldehyde. EGS/glycine treatment makes the zeta potential of synaptosomes more negative. Second, we modified glass coverslips with 3-aminopropyltriethoxysilane (APTES) to impart positive charges. EGS-fixed synaptosomes spontaneously attach to modified glasses via electrostatic attraction while maintaining good dispersion. Individual synaptic terminals are imaged by conventional fluorescence microscopy or by super-resolution techniques such as direct stochastic optical reconstruction microscopy (dSTORM). We examined tau protein by two-color and three-color dSTORM to understand its spatial distribution within mouse cortical synapses, observing tau colocalization with synaptic vesicles as well postsynaptic densities.Ushashi BhattacharyaJia-Fong JhouYi-Fong ZouGerald AbrigoShu-Wei LinYun-Hsuan ChenFan-Ching ChienHwan-Ching TaiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ushashi Bhattacharya Jia-Fong Jhou Yi-Fong Zou Gerald Abrigo Shu-Wei Lin Yun-Hsuan Chen Fan-Ching Chien Hwan-Ching Tai Surface charge manipulation and electrostatic immobilization of synaptosomes for super-resolution imaging: a study on tau compartmentalization
description	Abstract Synaptosomes are subcellular fractions prepared from brain tissues that are enriched in synaptic terminals, widely used for the study of neural transmission and synaptic dysfunction. Immunofluorescence imaging is increasingly applied to synaptosomes to investigate protein localization. However, conventional methods for imaging synaptosomes over glass coverslips suffer from formaldehyde-induced aggregation. Here, we developed a facile strategy to capture and image synaptosomes without aggregation artefacts. First, ethylene glycol bis(succinimidyl succinate) (EGS) is chosen as the chemical fixative to replace formaldehyde. EGS/glycine treatment makes the zeta potential of synaptosomes more negative. Second, we modified glass coverslips with 3-aminopropyltriethoxysilane (APTES) to impart positive charges. EGS-fixed synaptosomes spontaneously attach to modified glasses via electrostatic attraction while maintaining good dispersion. Individual synaptic terminals are imaged by conventional fluorescence microscopy or by super-resolution techniques such as direct stochastic optical reconstruction microscopy (dSTORM). We examined tau protein by two-color and three-color dSTORM to understand its spatial distribution within mouse cortical synapses, observing tau colocalization with synaptic vesicles as well postsynaptic densities.
format	article
author	Ushashi Bhattacharya Jia-Fong Jhou Yi-Fong Zou Gerald Abrigo Shu-Wei Lin Yun-Hsuan Chen Fan-Ching Chien Hwan-Ching Tai
author_facet	Ushashi Bhattacharya Jia-Fong Jhou Yi-Fong Zou Gerald Abrigo Shu-Wei Lin Yun-Hsuan Chen Fan-Ching Chien Hwan-Ching Tai
author_sort	Ushashi Bhattacharya
title	Surface charge manipulation and electrostatic immobilization of synaptosomes for super-resolution imaging: a study on tau compartmentalization
title_short	Surface charge manipulation and electrostatic immobilization of synaptosomes for super-resolution imaging: a study on tau compartmentalization
title_full	Surface charge manipulation and electrostatic immobilization of synaptosomes for super-resolution imaging: a study on tau compartmentalization
title_fullStr	Surface charge manipulation and electrostatic immobilization of synaptosomes for super-resolution imaging: a study on tau compartmentalization
title_full_unstemmed	Surface charge manipulation and electrostatic immobilization of synaptosomes for super-resolution imaging: a study on tau compartmentalization
title_sort	surface charge manipulation and electrostatic immobilization of synaptosomes for super-resolution imaging: a study on tau compartmentalization
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/dcce73005ca6481c969d0e8d56ddfc6b
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Surface charge manipulation and electrostatic immobilization of synaptosomes for super-resolution imaging: a study on tau compartmentalization

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