ArticleJournal of neuroscience methods2020
Biolistic transfection and expression analysis of acute cortical slices.
Article in Journal of neuroscience methods, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.
What it found
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Who cites it
3 citing papers in PubMed, 4 citations in OpenAlex.
- Reelin differentially shapes dendrite morphology of medial entorhinal cortical ocean and island cells.Development (Cambridge, England) · 2024Article
- Reelin Regulates Developmental Desynchronization Transition of Neocortical Network Activity.Biomolecules · 2024Article
- Reelin restricts dendritic growth of interneurons in the neocortex.Development (Cambridge, England) · 2021Article
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Authors and funding
10 authors at 3 institutions in 2 countries.
Funding
Abstract
backgroundBiolistic gene gun transfection has been used to transfect organotypic cultures (OTCs) or dissociated cultures in vitro. Here, we modified this technique to allow successful transfection of acute brain slices, followed by measurement of neuronal activity within a few hours. NEW
methodWe established biolistic transfection of murine acute cortical slices to measure calcium signals. Acute slices are mounted on plasma/thrombin coagulate and transfected with a calcium sensor. Imaging can be performed within 4 h post transfection without affecting cell viability.
resultsFour hours after GCaMP6s transfection, acute slices display remarkable fluorescent protein expression level allowing to study spontaneous activity and receptor pharmacology. While optimal gas pressure (150 psi) and gold particle size used (1 μm) confirm previously published protocols, the amount of 5 μg DNA was found to be optimal for particle coating. COMPARISON WITH EXISTING
methodsThe major advantage of this technique is the rapid disposition of acute slices for calcium imaging. No transgenic GECI expressing animals or OTC for long periods are required. In acute slices, network interaction and connectivity are preserved. The method allows to obtain physiological readouts within 4 h, before functional tissue modifications might come into effect. Limitations of this technique are random transfection, low expression efficiency when using specific promotors, and preclusion or genetic manipulations that require a prolonged time before physiological changes become measurable, such as expression of recombinant proteins that require transport to distant subcellular localizations.
conclusionThe method is optimal for short-time investigation of calcium signals in acute slices.
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