GSM4296225: RNA181102RH_26016_NAcCore_S4; Macaca mulatta; RNA-Seq
After the 12-month open access period, and without imposed abstinence, a previously described, detailed necropsy protocol (Daunais et al., 2010) was used to systematically collect tissues from all subjects. Briefly, monkeys were anesthetized with ketamine (10 mg kg − 1), maintained on isoflurane and perfused with ice-cold oxygenated monkey perfusion solution (containing [in mM] 124 NaCl, 23 NaHCO3, 3 NaH2PO4, 5 KCl, 2 MgSO4, 10 D-glucose, 2 CaCl2). Brains were quickly removed and sectioned along the coronal plane using a brain matrix (Daunais et al., 2010).The block containing the NAcc was initially selected by each individual's magnetic resonance imaging and verified using visible landmarks. In macaques, the NAcc is ~ 2 mm × 2 mm and extends ~ 3 mm rostral/caudal (Daunais et al., 2010). The core is differentiated from the shell based on visible landmarks such as the curvature of the internal capsule in which the area just ventral to its end is the NAcc. From the frozen 4 mm coronal brain block maintained on dry ice, a small circular dissection of ~ 1 mm^3 was made, taking care to not collect white matter from the tract (dorsal to the core). This relatively small dissection avoids the NAc shell and yields enough tissue for nucleic acid isolation. RNA was extracted from the NAcc using the All Prep DNA/RNA/microRNA Universal Kit (Qiagen Sciences, Germantown, MD, USA) following the manufacturer's recommendations. RNA libraries were prepared from cDNA for sequencing using Illumina TruSeq DNA library kit. Samples were multiplexed in 8 lanes with 3 samples per lane on an Illumina HiSeq 2500 sequencer. RNA-Seq type was strand-specific paired-end with PolyA+ strand-select protocol