This procedure was based on previous studies [9 (link), 81 , 82 (link)]. Rats were habituated to experimenter handling and the open field arena. Rats were fasted 16 h before the experiment. On the day of the experiment, rats received a continuous i.v. infusion of vehicle (buffered saline), (S)-ketamine (10 mg/kg), or HNK (10 mg/kg) over 40 min in an open field arena. Ten minutes after start of infusion, rats were injected (IP) with 13 MBq of 2-deoxy-2-[18F]-fluoro-D-glucose (FDG; Cardinal Health). After 30 min, rats were anesthetized with 1.5% isoflurane, placed on a custom-made bed of a nanoScan small animal PET/CT scanner (Mediso Medical Imaging Systems) and scanned for 20 min on a static acquisition protocol, followed by a CT scan. The PET data were reconstructed and coregistered to an MRI template using PMOD4.2 software environment (PMOD Technologies, Switzerland). The standardized uptake value ratio (SUVRWB) images were calculated using the whole brain as a reference region. All statistical parametric mapping analyses were performed using Matlab R2016 (Mathworks) and SPM12 (University College London). The ROI-based data was obtained using PMOD and the Schiffer rat VOI-atlas[83 (link)] and the statistical analyses were performed using GraphPad’s Prism 9.
Protocol detail
PET Imaging of Ketamine and Metabolite Effects
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Corresponding Organization : Johns Hopkins University
Other organizations : National Institute on Drug Abuse, National Institute of Neurological Disorders and Stroke, National Center for Advancing Translational Sciences, National Institute on Aging, National Institutes of Health, National Institute of Mental Health, University of Cyprus, VA Maryland Health Care System
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Variable analysis
independent variables
- Infusion treatment (vehicle, (S)-ketamine, or HNK)
- Standardized uptake value ratio (SUVRWB) in the brain
- Rat habituation to experimenter handling and open field arena
- Rat fasting duration (16 h)
- FDG injection timing (10 min after start of infusion)
- Anesthesia and scanning protocol (1.5% isoflurane, 20 min static PET/CT scan)
- Data analysis (PET data reconstruction, coregistration to MRI template, SUVRWB calculation, statistical parametric mapping, ROI-based analysis)
- Vehicle (buffered saline) infusion
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