An Improved Method for Growing Primary Neurons on Electron Microscopy Grids Co-Cultured with Astrocytes

Ishika Kumar; Anju Paudyal; Anna Kádková; Michelle Stewart; Jakob Balslev Sørensen; Julika Radecke

doi:10.3390/ijms242015191

An Improved Method for Growing Primary Neurons on Electron Microscopy Grids Co-Cultured with Astrocytes

Ishika Kumar, Anju Paudyal, Anna Kádková, Michelle Stewart, Jakob Balslev Sørensen, Julika Radecke^*

^*Corresponding author af dette arbejde

Neuronal Signalling

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

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Abstract

With the increasing popularity of cryo-electron tomography (cryo-ET) in recent years, the quest to establish a method for growing primary neurons directly on electron microscopy grids (EM grids) has been ongoing. Here we describe a straightforward way to establish a mature neuronal network on EM grids, which includes formation of synaptic contacts. These synapses were thin enough to allow for direct visualization of small filaments such as SNARE proteins tethering the synaptic vesicle (SV) to the active zone plasma membrane on a Titan Krios without prior focused ion-beam milling.

Originalsprog	Engelsk
Artikelnummer	15191
Tidsskrift	International Journal of Molecular Sciences
Vol/bind	24
Udgave nummer	20
Antal sider	15
ISSN	1661-6596
DOI	https://doi.org/10.3390/ijms242015191
Status	Udgivet - 2023

Bibliografisk note

Funding Information:
We thank the Novo Nordisk Foundation (NNF17OC0028516 to JBS) and the Carlsberg Foundation (CF17-0875 to JBS) for support. We acknowledge Diamond for access and support of the cryo-EM facilities at the UK national electron Bio-Imaging Centre (eBIC), proposal NT29869, funded by the Wellcome Trust, MRC and BBSRC. Work at the Mary Lyon Centre was underpinned by funding from the Medical Research Council grant A410.

Publisher Copyright:
© 2023 by the authors.

Adgang til dokumentet

10.3390/ijms242015191Licens: CC BY

FulltextForlagets udgivne version, 12,2 MBLicens: CC BY

Citationsformater

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AU - Radecke, Julika

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