Butler-Ryan, R and Wood, IC orcid.org/0000-0003-1886-2533 (2021) Efficient infection of organotypic hippocampal slice cultures with adenovirus carrying the transgene REST/NRSF. Journal of Neuroscience Methods, 356. 109147. ISSN 0165-0270
Abstract
Organotypic hippocampal slice cultures provide a useful platform maintaining hippocampal structure and synaptic connections of the brain over weeks in culture with ease of in vitro manipulations. Gene transfer is a particularly desirable tool for using with them but current difficulties with transformation of transgenes into these cultures is a barrier to their use in research. Previous quantifications of viral infections have shown low transformation rates and have relied upon invasive microinjections. In this paper we present an efficient way of infecting organotypic cultures with adenovirus at the acute slice stage that does not require injection. We use the adenoviral delivery system to introduce the transcription factor REST and a GFP marker, providing around 41 % cellular infection spread throughout the entire slice culture and promoting transgene expression for weeks in vitro. GFP expression was observed most intensely in the slices when they were infected just a few hours after plating and was shown to infect neurons and microglia. We decided to use the transcription factor REST/NRSF as an example transgene which was delivered into cells via the adenoviral construct, conferring overexpression of REST in addition to the GFP marker. This outlines a technique whereby adenoviral infection of organotypic cultures can infect neurons with good efficiency and confer successful manipulation of genetic factors within the cell.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | Crown Copyright © 2021. Published by Elsevier B.V. All rights reserved. This is an author produced version of an article published in Journal of Neuroscience Methods. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | Organotypic; Hippocampus; Adenovirus; REST; NRSF |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biomedical Sciences (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 23 Mar 2021 11:32 |
Last Modified: | 23 Mar 2022 01:38 |
Status: | Published |
Publisher: | Elsevier |
Identification Number: | 10.1016/j.jneumeth.2021.109147 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:172463 |