Silencing Noggin in Mouse Hippocampus

Direct hippocampal injections of lentivirus expressing a shRNA against noggin (Bond et al., 2014 (link); Brooker et al., 2017 (link)) were performed using a mouse stereotaxic instrument (Stoelting Co), Quintessential Stereotaxic Injector (Stoelting Co), and a Hamilton microsyringe (5 μl, NEUROS Model, 33 g, Blunt). Mice were anesthetized via inhalation of isoflurane. Two small craniotomies were performed over the hippocampus of each hemisphere (coordinates relative to bregma: 2 mm posterior, 1.5 mm lateral, 1.9 mm ventral) and 2 μl of virus was injected bilaterally into the DG at a rate of 0.5 μl/min. Two weeks postinjection, levels of noggin protein were analyzed, and any mice lacking successful viral infection in the DG were excluded from the study.