At 29 (replicate 1) and 31 (replicates 2 and 3) days of age, piglets were subjected to magnetic resonance imaging (MRI) procedures. Scanning procedures were implemented on a Siemens MAGNETOM Trio 3 T Imager using a 32-channel head coil. Upon arrival to the Biomedical Imaging Center located in the Beckman Institute for Advanced Science and Technology, piglets were anesthetized via intramuscular injection of Telazol (0.07 mg/kg body weight; Zoetis, Florham Park, NJ, USA). Once sedated, piglets were transferred to the MRI scanner and maintained on 2% isoflurane/98% oxygen (Isoflurane, Piramal Healthcare, Bethlehem, PA, USA) for the entirety of the 60-min scan. Piglet vital signs were monitored throughout the scan using an MRI-compatible pulse oximeter, and upon completion of the scan, piglets were maintained under anesthesia and immediately euthanized for tissue collection. Detailed methods for manual brain segmentation, volumetric assessment, voxel-based morphometry, and diffusion tensor imaging were previously described (15 (link), 16 (link)).
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