Cultures were whole cell patch clamped as described previously. Nicotine was perfused in extracellular medium in increasing amounts 0.1 μM to 6 μM in 3 minute intervals. EPSC frequencies were counted using the pClampfit software (Molecular dynamics). Frequencies were normalized to the first 30 seconds of wash buffer and dosage frequencies were averaged over 30 second intervals within first minute. Amplitudes within each 10 second interval were also averaged in the same time frame.
