Single-Cell Electroporation across Different Organotypic Slice Culture of Mouse Hippocampal Excitatory and Class-Specific Inhibitory Neurons
Student Authors
Amy CheungDavid G. Keener
Academic Program
MD/PhDUMass Chan Affiliations
Graduate School of Biomedical SciencesBrudnick Neuropsychiatric Research Institute
Neurobiology
Futai Lab
Document Type
Journal ArticlePublication Date
2020-10-06Keywords
Investigative TechniquesLaboratory and Basic Science Research
Molecular Biology
Neuroscience and Neurobiology
Metadata
Show full item recordAbstract
Electroporation has established itself as a critical method for transferring specific genes into cells to understand their function. Here, we describe a single-cell electroporation technique that maximizes the efficiency (~80%) of in vitro gene transfection in excitatory and class-specific inhibitory neurons in mouse organotypic hippocampal slice culture. Using large glass electrodes, tetrodotoxin-containing artificial cerebrospinal fluid and mild electrical pulses, we delivered a gene of interest into cultured hippocampal CA1 pyramidal neurons and inhibitory interneurons. Moreover, electroporation could be carried out in cultured hippocampal slices up to 21 days in vitro with no reduction in transfection efficiency, allowing for the study of varying slice culture developmental stages. With interest growing in examining the molecular functions of genes across a diverse range of cell types, our method demonstrates a reliable and straightforward approach to in vitro gene transfection in mouse brain tissue that can be performed with existing electrophysiology equipment and techniques.Source
Keener DG, Cheung A, Futai K. Single-Cell Electroporation across Different Organotypic Slice Culture of Mouse Hippocampal Excitatory and Class-Specific Inhibitory Neurons. J Vis Exp. 2020 Oct 6;(164). doi: 10.3791/61662. PMID: 33104060. Link to article on publisher's site
DOI
10.3791/61662Permanent Link to this Item
http://hdl.handle.net/20.500.14038/29611PubMed ID
33104060Related Resources
ae974a485f413a2113503eed53cd6c53
10.3791/61662