Viral-Mediated Gene Transfer to Study the Behavioral Correlates of CREB Function in the Nucleus Accumbens of Rats - Drugs of Abuse: Neurological Reviews and Protocols - page 326

2.4. Surgery

1. Rats, male, 300-325 g at the time of surgery.

2. Stereotaxic instrument (two arms: left and right).

L) with 26-gauge, blunt-tipped needles.
4. Sterile fi eld (if aseptic procedures required).
5. Distilled water, and 95% ethanol.
6. 0.9% Saline.
7. Isopropyl alcohol and Betadine wipes.
8. Standard surgical instruments (scalpel, hemostats, autoclip applier, etc.).
9. Anesthetic (if pentobarbital, then also atropine).
10. Drill.
11. Timer.

3. Two Hamilton syringes (5 µ

2.5. Histology
2.5.1. Fixed Tissue

1. Peristaltic perfusion pump.

2. Physiological saline (~500 mL per rat).

3. 4% Paraformaldehyde (~500 mL per rat).

4. Anesthetic.

5. Standard surgical instruments.

6. Decapitator.

2.5.2. Fresh Tissue

1. Decapitator.

2. Artifi cial cerebrospinal fl uid (aCSF) buffer: 126 m M NaCl, 5 m M KCl, 1.25 m M
NaH 2 PO 4 , 25 m M NaHCO 3 , 2 m M CaCl 2 , 2 m M MgCl 2 , and 10 m M D -glucose,
at pH 7.4.

3. Tissue slicer (Stoelting, 51425).

4. Dry ice.

5. Tissue punches.

6. Microcentrifuge tubes for sample collection.

2.6. Immunohistochemistry

1. Freezing microtome.

2. Normal goat serum (NGS).

3. Bovine serum albumin (BSA).

4. Rabbit polyclonal antibody to CREB, 1 1000 (Cell Signaling).

5. Goat biotinylated antibody to rabbit IgG, 1 200 (Vector Laboratories).

6. Avidin-biotin (ABC Elite, Vector).

7. 3,3 -Diaminobenzidine tetrahydrochloride (DAB).

8. Slides (Superfrost Plus, Fisher).

9. Dehydration/staining kit.