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2 with KOH. The patch electrode was controlled using a motorized micromanipulator (Sutter Instruments, Novato, CA). An Axopatch-1C amplifier (Axon Instruments, Foster City, CA) and pClamp9 software (Molecular Devices, Sunnyvale, CA) were used for recording. The junction potential was estimated at -15.2 mV and was corrected for. All drugs were obtained from Sigma-Aldrich (St. Louis, MO), unless otherwise noted. Intrinsic property characterization. Intrinsic properties were characterized in current-clamp mode following published protocols with minor modifications 11. A mixture of synaptic blockers was used to inhibit synaptic activity: 5 ��M 6,7-dinitroquinoxaline-2,3-dione disodium salt (DNQX), 5 ��M bicuculline and 25 ��M DL-2-amino-5-phosphonopentanoic acid sodium salt (DL-AP5) (Abcam, Toronto, Canada). Data analysis was done using custom Matlab software (MathWorks, Natick, MA). Once the whole-cell configuration was achieved, the cell��s Casein kinase 1 resting membrane potential was noted and its spontaneous firing, if any, was recorded for 30 s. Access resistance and resting membrane potential were checked at regular intervals (every ~5 min) throughout the recording of the cell. The frequency-current (f-I) profiles of the pyramidal cells are important to characterize, as we aim for our single cell model to respond to a variety of synaptic input strengths with frequencies similar to that observed experimentally. These f-I curves were determined by applying depolarizing current steps of 1 s duration to cells held in current clamp. Amplitudes were increased incrementally with step sizes of 25 pA CFTR inhibitor manufacturer for one of four cells, and 10 pA for three of four cells. The initial firing frequency was determined based on the inverse of the first inter-spike interval, and the final frequency was based on the inverse of the last inter-spike interval in the 1-second depolarizing step. For each cell, the approximate linear slope of the f-I curve above 5 Hz was determined using a least squares method. These values were chosen selleck products since above 5 Hz the slope was well-approximated by linearization. In addition, the minimum amount of current required to initiate a spike, the rheobase current ( I rheo in pA), was determined. The action potential threshold was set to be the first voltage point such that the slope of the membrane potential exceeded 20 mV/ms 12. The spike width was determined at the threshold value. In addition, the spike height from threshold and the minimum membrane potential reached following the spike were measured. Recordings were kept for analysis only if the neuron remained stationary; spikes overshot 0 mV (-15 mV junction potential corrected) and access resistance