Document Type
Article
Publication Date
10-18-2017
Publication Title
MethodsX
Volume
4
Abstract
Understanding how neuromodulators influence synaptic transmission and intrinsic excitability within the entorhinal cortex (EC) is critical to furthering our understanding of the molecular and cellular aspects of this region. Organotypic cultures can provide a cost-effective means to employ selective molecular biological strategies in elucidating cellular mechanisms of neuromodulation in the EC. We therefore adapted our acute slice model for organotypic culture applications and optimized a protocol for the preparation and biolistic transfection of cultured horizontal EC slices. Here, we present our detailed protocol for culturing EC slices. Using an n-methyl-D-glucamine (NMDG)-containing cutting solution, we obtain healthy EC slice cultures for electrophysiological recordings. We also present our protocol for the preparation of “bullets” carrying one or more constructs and demonstrate successful transfection of EC slices. We build upon previous methods and highlight specific aspects in our method that greatly improved the quality of our results. We validate our methods using immunohistochemical, imaging, and electrophysiological techniques. The novelty of this method is that it provides a description of culturing and transfection of EC neurons for specifically addressing their functionality. This method will enable researchers interested in entorhinal function to quickly adopt a similar slice culture transfection system for their own investigations.
First Page
360
Last Page
371
DOI
10.1016/j.mex.2017.10.003
ISSN
22150161
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Recommended Citation
Nicholas I. Cilz, James E. Porter, and Saobo Lei. "A Protocol for Preparation and Transfection of Rat Entorhinal Cortex Organotypic Cultures for Electrophysiological Whole-Cell Recordings" (2017). Biomedical Sciences Faculty Publications. 18.
https://commons.und.edu/bms-fac/18