Mice were placed on doxycycline chow 24 hr prior to surgery (40 mg/kg, Bio-Serv, Flemington, NJ). On the day of surgery, mice were anaesthetized using isoflurane (3% induction, 1.5% maintenance during surgery) and secured to a stereotactic frame (Kopf) with a heating pad to maintain body temperature. Mice were injected sub-dermally with Buprenex (1 mg/kg) and given topical Lidocaine (20 mg/mL) for analgesia (Hi-Tech Pharmacal, Amityville, NY). Following exposure of the skull, a small craniotomy was made overlying the target brain structure. Using a glass pipette (50 µm tip) connected to a Nanoject II (Drummond Scientific, Broomall, PA) injector, AAVs were delivered (50 nL/infusion, 30 sec between infusions) to the brain, and allowed to diffuse for 10 min before withdrawing the pipette. The coordinates of the target brain structures in reference to bregma (mm) and injection volumes (nL) were as follows: dentate gyrus (AP: −1.9, ML: ± 1.4, DV: −2.05; 600), dorsal CA3 (AP: −1.9, ML: ± 1.85, DV: −2.1; 500), basolateral amygdala (AP: −1.6, ML: ± 3.25, DV: −3.6 from pial surface; 1000), and central amygdala (AP: −1.4, ML: ± 2.85, DV: −3.7 from pial surface; 300). All injections were performed bilaterally. Mice were kept on doxycycline chow and allowed to recover for 7–14 days following surgery.
Rats were anesthetized with isoflurane. A midline incision was made to expose the skull, and a small craniotomy was made unilaterally above the medial prefrontal cortex (mPFC). Recombinant AAV was injected into the prelimbic cortex (PL) of the mPFC. A stainless steel needle with the beveled tip facing laterally (31 gauge; Hamilton Company, Reno, Nevada) was directed to the PL (AP: +2.5, ML: ±0.5, DV: −2.0 from pial surface). For the CRAM experiment in rats, CAV2-Cre was bilaterally injected to the dorsal medial striatum (AP: +0.1, ML: ± 2.0, DV: −3.5 from pial surface). Virus (1.0 μL/hemisphere) was infused over 10 min (0.1 μL/min), followed by an additional 10 min to allow diffusion of the virus from the needle tip. The skin was sealed with Vetbond tissue adhesive.
Rats were anesthetized with isoflurane. A midline incision was made to expose the skull, and a small craniotomy was made unilaterally above the medial prefrontal cortex (mPFC). Recombinant AAV was injected into the prelimbic cortex (PL) of the mPFC. A stainless steel needle with the beveled tip facing laterally (31 gauge; Hamilton Company, Reno, Nevada) was directed to the PL (AP: +2.5, ML: ±0.5, DV: −2.0 from pial surface). For the CRAM experiment in rats, CAV2-Cre was bilaterally injected to the dorsal medial striatum (AP: +0.1, ML: ± 2.0, DV: −3.5 from pial surface). Virus (1.0 μL/hemisphere) was infused over 10 min (0.1 μL/min), followed by an additional 10 min to allow diffusion of the virus from the needle tip. The skin was sealed with Vetbond tissue adhesive.
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