Amygdalar ASIC1a Modulation in Mice

For the cannula placement procedure, mice were anesthetized with isoflurane through an anesthetic vaporizer, secured to the stereotaxic instrument, and the cannula made from a 25-gauge needle was inserted bilaterally into LA and basolateral amygdala (relative to bregma: –1.2 mm anteroposterior; ±3.5 mm mediolateral; –4.3 mm dorsoventral) (16 (link), 36 (link)). Dental cement secured the cannula and bone anchor screw in place. Mice recovered for 4–5 days before any subsequent testing was carried out. A 10 μL Hamilton syringe connected to a 30-gauge injector was inserted 1 mm past the cannula tip to inject the selective ASIC1a inhibitor, PcTX-1 (100 nM, Allomone Labs) or anisomycin (62.5 μg/μL, Cayman Chemical). The chemicals were diluted in 1 μL artificial cerebrospinal fluid (ACSF), pH 7.3, and injected over 5 minutes each side. The injection sites were mapped postmortem by sectioning the brain (10 μm coronal) and examining Cresyl violet staining using a Nissl Stain Kit (FD Neuro Technologies). Only animals that had a correctly placed cannula in the amygdala were included in the statistical analysis.