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To test the effects of stimulation intensity, 4 pigs from the STN DBS group and 3 pigs from the EN/GPi DBS group were randomly selected to receive biphasic 2?V pulses at 130?Hz and pulse widths of 500?��s followed by a ~?10?min rest after the first fMRI scan. For the single subject in which SN was the target, stimulation parameters of biphasic 300?��A pulses at 60?Hz, and pulse widths of 2?ms were applied. Upon completion of the experimental fMRI DBS procedure, subjects were deeply anesthetized with intravenous sodium pentobarbital (100?mg/kg) and euthanized. The brain was extracted and kept in 10% formalin for two weeks. The brain was then sectioned at 250?��m in the coronal plane by a vibratome (VT 1000?s, Leica) to confirm the DBS lead location by see more visual inspection of the DBS lead track. The electrode position was marked in the pig brain atlas as shown in Fig.?1B. The fMRI data were converted MEK inhibitor into BrainVoyager data format. A standard sequence of post-processing steps, including 3D motion correction and temporal filtering (Gaussian filter; FWHM 3 data points), was implemented in the BrainVoyager QX software and applied to each data set. Functional activation was analyzed by correlating the observed signal intensity changes in each voxel with the given stimulus protocol. Based on the results of this procedure, an appropriate activation map was generated for each subject. To account for hemodynamic delay, the stimulus representing the block design was convolved with a double-gamma hemodynamic response function (onset 0?s, time to response peak 5?s, time to undershoot peak 15?s). To correct for multiple comparisons and exclude false positive voxels, we considered only voxels with a significance level less than the False Discovery Rate (FDR)?crotamiton on both 2-dimensional image leaving only the brain by Analyzer 10.0 software (Mayo Clinic) and then the image was used for a full affine (12 parameters) normalization to the 3D atlas by BrainVoyager QX. Event-related BOLD responses were calculated by measuring the signal intensities that began 10 volumes before the stimulus onset and continued for 25 volumes after the stimulus ended. The averaged signal intensity within the appropriate area in the first 10 volumes was set to 100% signal intensity as a baseline, and post-stimulus signal variation was calculated relative to it.