Male C57Bl/6J mouse 12 weeks old at the time of sacrifice
Mice were briefly anesthetized with isoflurane then rapidly decapitated. Brains were submerged in oxygenated (95% O2-5% CO2) ice-cold sucrose-artificial cerebrospinal fluid solution (in mM: 194 sucrose, 20 NaCl, 4.4 KCl, 2 CaCl2, 1 MgCl2, 1.2 NaH2PO4, 10.0 glucose, and 26.0 NaHCO3) and coronal brain slices (300 um) were made using a vibratome (Leica). Slices were transferred onto a glass Petri dish with a transfer pipette and excess fluid was drained from the area surrounding the tissue. The Petri dish was placed on top of a chilled aluminum block and tissue was brought to a semi-frozen state before punches were taken.
Stat-60 (Tel-Test) extraction of total RNA was performed according to the manufacturer's instructions.
Starting with 10 ng of total RNA, cRNA targets were generated using the two-cycle target labeling method.
Labeled cRNA was purified, and 20 ?g of cRNA was fragmented to a range of 35 to 200 bases in length. Samples were hybridized at 450C for 16 h to Affymetrix mouse430_2 chips GeneChips were washed and stained in the Affymetrix Fluidic
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000
Gene expression data from nucleus accumbens of mouse
The data were analyzed with dChip softwareusing invariant-set normalization method.