Brain slices were placed in an interface chamber (32–34°C), oxygenated with 95% O2–5% CO2, and perfused with aCSF containing 100 μM of 4-aminopyridine (4-AP) to induce epileptiform activity.30 (link),31 (link) Glass electrodes filled with aCSF were placed in the neocortex (motor and somatosensory regions, up to the most anterior aspect of the hippocampus, layer IV/V) and visualized with a stereomicroscope (AmScope). Extracellular field potentials were recorded using a low noise differential amplifier (100x gain, DP-311, Warner Instruments) and digitized at 2 kHz (IX/408, iWorx Systems Incorporated). Epileptiform activity analysis was measured using a custom-written macro in IgorPro v8.04 (WaveMetrics).13 (link),32 (link) Events lasting less than 10 seconds were considered interictals, and longer events were considered seizure-like events (ictal)33 (link).
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Inducing Epileptiform Activity in Cortical Slices
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Corresponding Organization : University of Iowa
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Variable analysis
independent variables
- 4-aminopyridine (4-AP) concentration (100 μM)
- Extracellular field potentials
- Epileptiform activity
- Temperature (32–34°C)
- Oxygenation (95% O2–5% CO2)
- Perfusion with artificial cerebrospinal fluid (aCSF)
- No positive or negative controls were explicitly mentioned.
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