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A versatile tool for the analysis of neuronal survival

Authors
  • Mergenthaler, Philipp
  • Wendland, Kristin
  • Meisel, Andreas1, 2, 3, 4, 5
  • 1 Department of Experimental Neurology
  • 2 Department of Neurology
  • 3 Center for Stroke Research
  • 4 NeuroCure Cluster of Excellence
  • 5 Charité University Medicine Berlin
Type
Published Article
Journal
Methods
Publication Date
Jan 01, 2013
Volume
66
Issue
2
Pages
325–329
Identifiers
DOI: 10.1016/j.tins.2013.07.001
Source
Elsevier
Keywords
License
Unknown

Abstract

To understand the principles that govern mechanisms of neuronal survival or death it is necessary to systematically model these processes. Methods involving overexpression or knockdown of a gene of interest using non-viral transfection of primary neurons can easily be adapted to study cell death pathways in primary neurons. However, common biochemical approaches to measure cell death are insufficient to measure neuronal viability in these systems. To investigate the functional role of genes in cultured neurons, we therefore established a cell-based assay using a cotransfection/cocultivation approach in primary cortical neurons from mouse or rat. Using this method, it is possible to use well-established cell culture models of neuronal damage, and to analyze cell survival in genetically different neurons on a single-cell basis following apoptotic stimuli under identical conditions. The duration of the entire protocol is 10days. Finally, the method may be applicable to a wide range of damage models, primary cells, and cell lines as well as it can be used for high content screening (HCS) studies and downstream image cytometry.

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