2.8.7. Sacrifice and brain processing

After completion of the behavioral experiments, rats were fasted for 12 h during the light phase and at dark onset. They received an injection of amylin (50 μg/kg i.p.) or a vehicle. Ninety min later, animals were deeply anesthetized, perfused, and processed as above. Brains of LPBN (bregma level −8.72 mm to −10.04 mm) and CeA (bregma level −1.60 mm to −3.30 mm) were cut at the cryostat. Sections were immunostained for CGRP/Fos to verify the CGRP depletion in LPBN and neural activation through the LPBN and the LPBN projection site, the CeA.