Figure 3
From: GSK-3β activation is required for ZIP-induced disruption of learned fear
Pre-injection of BIO-acetoxime or GSK-3β shRNA attenuates ZIP-induced memory disruption. (a) Schematic of experimental design for experiments shown in (a)–(c). Rats were fear-conditioned on Day 1. BIO-acetoxime (or vehicle) was microinjected 1 h before the ZIP (or vehicle) injection on Day 3. ZIP (10 nmol/µL/hemisphere) or BIO-acetoxime (75 pmol/0.5 µL/hemisphere) was injected into the LA. The memory retention test was performed on Day 4. Schematic illustrations of injection target are displayed in Fig. S2a and b (b) Pre-injection of BIO-acetoxime attenuated ZIP-induced memory disruption. Veh-Veh (vehicle of BIO-acetoxime for 1st injection, vehicle of ZIP for 2nd injection), Veh-ZIP (vehicle of BIO-acetoxime for 1st injection, ZIP for 2nd injection), Bio-Veh (BIO-acetoxime for 1st injection, vehicle of ZIP for 2nd injection), Bio-ZIP (BIO-acetoxime for 1st injection, ZIP for 2nd injection). (c) Open field test was performed for all experimental groups. A new set of rats were used for the open field test. No significant difference was observed between the groups. (d) Schematic of experimental design for experiments shown in (d)–(f). Lentiviral particles containing GSK-3β shRNA or control shRNA were injected into the LA 24 h before fear conditioning. Vehicle or ZIP was injected 48 h after fear conditioning, and the memory retention test or the preparation of LA synaptosomes was performed on the following day. Schematic illustrations of injection target are displayed in Fig. S2c. (e) Synaptosomal GSK-3β (left) but not synaptosomal actin (right) was reduced by microinjection of GSK-3β shRNA. Full-length blots are displayed in Fig. S3c–f. (F) Pre-injection of GSK-3β shRNA attenuated ZIP-induced memory disruption. Ctrl shRNA-Veh (Ctrl shRNA for 1st injection, vehicle of ZIP for 2nd injection), Ctrl shRNA-ZIP (Ctrl shRNA for 1st injection, ZIP for 2nd injection), GSK-3β shRNA-Veh (GSK-3β shRNA for 1st injection, vehicle of ZIP for 2nd injection), GSK-3β shRNA-ZIP (GSK-3β shRNA for 1st injection, ZIP for 2nd injection). *p < 0.05.
