Affymetrix mouse gene 1.1 st array plates

Laser microdissection was performed using a Leica LMD6500 system (40X objective in the fluorescence mode for SST neurons or bright-field mode for pyramidal neurons). Cutting and collection steps were subsequently examined in fluorescent or bright-field mode. 100 cells from the mouse cingulate cortex per animal were collected in 0.5 ml microtube caps (Ambion, Foster City, CA, USA) and lysed by vortexing for 30 sec in 150 μl of RLT Buffer Plus (Qiagen, Valencia, CA, USA) within a 3-hour time span, and stored at 80 °C until further processing. Total RNA was then extracted using the RNeasy Plus Micro Kit (Qiagen) according to the manufacturer’s instructions. RNA samples were amplified with NuGEN® Ovation Pico WTA System V2 (NuGen, San Carlos, CA). The fragmented labeled cDNA samples were processed and hybridized to Affymetrix® Mouse Gene 1.1 ST Array Plates (Affymetrix). Gene expression information was obtained with the Affymetrix console software. Genes showing significant differences (Student t-test p<0.005; >1.2 fold cutoff) in expression between SST neurons from stressed and unstressed mice were analyzed using the IPA software (Ingenuity Systems, Redwood, CA).