Mouse hippocampal neurons were isolated and cultured from floxed-GABABR1 mice as described previously.27 (link),46 (link) Cells were cultured in poly-d -lysine-coated (0.1 mg/mL) plastic dishes and/or glass coverslips in a neuron maintenance medium27 (link),46 (link) for 3 days and supplemented with cytosine β-d -arabinofuranoside (3 μmol/L; Sigma, St. Louis, MO, USA) for an additional 3–4 days. These neurons were then infected with adenoviruses (8 pfu/cell) carrying a cDNA encoding Cre recombinase (Ad-Cre; Microbix Inc., Toronto, Canada) or empty viral vector (Ad-Cont Microbix Inc., Toronto, Canada) as described previously.27 (link),46 (link)
Protocol detail
Conditional Knockout of Hippocampal GABA Receptors
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Corresponding Organization : University of California, San Francisco
Other organizations : Neurological Surgery
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Variable analysis
independent variables
- Presence of Cre recombinase (Ad-Cre) or empty viral vector (Ad-Cont)
- Not explicitly mentioned
- Culture of mouse hippocampal neurons in poly-D-lysine-coated plastic dishes and/or glass coverslips
- Neuron maintenance medium for 3 days
- Supplementation with cytosine β-D-arabinofuranoside (3 μmol/L) for an additional 3-4 days
- Positive control: Cells infected with adenovirus carrying Cre recombinase (Ad-Cre)
- Negative control: Cells infected with empty viral vector (Ad-Cont)
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