GSM6944210: sustained CeA male MR1 79_CeA; Mus musculus; RNA-Seq
Mouse brains were snap-frozen on dry ice and stored at -80°C. Regions of interest (ACC, BNST, BLA, and CeA) were isolated by a brain punching technique from the whole-mount brain in the cryostat Leica CM3050 S (Leica Biosystems, Nussloch, Germany) at -20°C using biopsy pens EMS Rapid-Core (Electron Microscopy Sciences, Hatfield, USA). To localize the subregions of interest in the brain, punching was carried out by referencing toluidine blue staining of brain slices (Franklin & Paxinos, 2007).  ACC was sampled approximately from 1.34 to 0.5mm (from bregma) using a 1mm pen. BNST (0.5: -0.22)mm, BLA (-0.94: -2.06)mm, and CeA (-0.94: -1.82)mm were sampled bilaterally using a 0.5mm pen. Biopsies were stored at -80°C for further RNA extraction. RNA was harvested using Rneasy mini plus kit (Qiagen). 1.3 ug of total RNA was used for the construction of sequencing libraries. All steps were carried out on ice. Tissue biopsies were moved to precooled Precellys tubes containing ice-cold ceramic beads for homogenization via Precellys 24 (Peqlab, Erlangen, Germany) (600 g, 20 s). Extraction was performed using RNeasy® mini kit (Qiagen, Hilden, Germany) according to the protocol provided by the manufacturer.