Monkeys were trained to extend a leg and remain stationary during venipuncture through a process of reinforcing successive approximations with food treats. Once trained, blood samples were taken from the lateral saphenous vein and collected in silicone coated serum vacutainer tubes for serum and K3EDTA coated vacutainer tubes (BD Biosciences, Haryana, India), for plasma. Samples were collected prior to the morning dose of the antipsychotic and therefore represented trough levels. Blood in silica coated tubes was centrifuged at 1600 x g for 30 min. Serum was collected apportioned in 0.5 ml aliquots then stored at −80°C until assayed. Blood in K3EDTA coated tubes was centrifuged 1000–2000 x g for 10–15 min at 4°C, then apportioned in 0.5 ml aliquots and transferred to sterile polypropylene tubes. Aliquots used for blood chemistry analysis were stored at 4°C and shipped accordingly while aliquots for analysis of drug concentrations or biochemical assays were stored at −80°C until assayed.
After the 6-month treatment protocol, monkeys were sedated with ketamine followed by an overdose of sodium pentobarbital then transcardially perfused with ice cold phosphate buffered saline. Brains were removed and cut into four mm thick coronal slabs using a brain matrix (Ted Pella, Redding, CA, USA; Cat# 15039). Brain slabs were rapidly frozen on metal plates cooled by dry ice and stored at −80oC. Time from the beginning of transcardial perfusion to freezing of brain tissue was designated as the post-mortem interval (PMI) and ranged from 38 to 87 min and did not differ significantly between groups. Brain tissue was excised using a Miltex 3.5 mm diameter short handle biopsy punch. Care was taken to maximize consistency of the punch procedure across all brains. The ventral and dorsal banks of the principal sulcus were dissected as representative of DLPFC (Area 46). Brain tissue corresponding to the rostral and intermediate EC (Area 28) was dissected, immediately rostral to the appearance of the hippocampus, located between the rhinal fissure and the amygdala, according to anatomical landmarks and stereotaxic atlas (83 ).
After the 6-month treatment protocol, monkeys were sedated with ketamine followed by an overdose of sodium pentobarbital then transcardially perfused with ice cold phosphate buffered saline. Brains were removed and cut into four mm thick coronal slabs using a brain matrix (Ted Pella, Redding, CA, USA; Cat# 15039). Brain slabs were rapidly frozen on metal plates cooled by dry ice and stored at −80oC. Time from the beginning of transcardial perfusion to freezing of brain tissue was designated as the post-mortem interval (PMI) and ranged from 38 to 87 min and did not differ significantly between groups. Brain tissue was excised using a Miltex 3.5 mm diameter short handle biopsy punch. Care was taken to maximize consistency of the punch procedure across all brains. The ventral and dorsal banks of the principal sulcus were dissected as representative of DLPFC (Area 46). Brain tissue corresponding to the rostral and intermediate EC (Area 28) was dissected, immediately rostral to the appearance of the hippocampus, located between the rhinal fissure and the amygdala, according to anatomical landmarks and stereotaxic atlas (83 ).
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