Computational protocol: Staged anticonvulsant screening for chronic epilepsy

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Protocol publication

[…] Extracellular field potentials were recorded in the CA1 and/or CA3 pyramidal cell layer of intact hippocampi and organotypic hippocampal slices in a conventional submerged chamber using tungsten‐coated microelectrodes and ISO‐DAM8A amplifier (World Precision Instruments). Oxygenated (95% O2 and 5% CO2) artificial cerebrospinal fluid containing 126 mmol/L NaCl, 3.5 mmol/L KCl, 2 mmol/L CaCl2, 1.3 mmol/L MgCl2, 25 mmol/L NaHCO3, 1.2 mmol/L NaH2PO4, and 11 mmol/L glucose (pH 7.4), was continuously perfused at 33 ± 0.5°C. Flow rate was 2.5 mL/min. Before the actual recording, slices were allowed to stabilize in the recording chamber for 30–60 min. Electrical signals were digitized using an analog‐to‐digital converter (DigiData 1322A, Axon Instruments). pCLAMP 8.2 (Axon Instruments) and SigmaPlot 11.0 (Systat Software) programs were used for data acquisition and analysis. Recordings were sampled at 10 KHz and filtered from 1 Hz to 2 kHz. Ictal‐like epileptiform discharges were defined as hyper‐synchronous large amplitude (×3 baseline) and high‐frequency population spikes followed by sustained ictal‐tonic and intermittent ictal‐clonic after discharges, with the duration of the population spike and after‐discharge complex lasting more than 5–10 sec. Power spectrum analysis was performed from the electrical signals after applying a Hamming window function. […]

Pipeline specifications

Software tools pCLAMP, SigmaPlot
Applications Miscellaneous, Patch-clamp
Organisms Homo sapiens
Diseases Epilepsy
Chemicals Lactic Acid