Eyfp wpre hgh

Standard operating procedures for sterile surgery were employed. The AAV2.hSyn.ChR2(H134R)-eYFP.WPRE.hGH (#26973, Addgene USA) was diluted in PBS to a final concentration of 3.6 × 10⁹ vc/uL. Rats were anesthetized and maintained with isoflurane (5% for induction, 2.5% for maintenance), received saline (3 mL subcutaneously) for hydration, and Baytril antibiotic (5 mg/kg) and Xylocaine (7 mg/kg) local anesthetic applied around the surgical site. The rat was positioned on the stereotaxic frame, blood pressure monitored by pulse oximetry (MouseOx, STARR Life Sciences Corp, USA), and injection site body temperature maintained on a heating pad with rectal feedback probe (TC-1000 Temperature Controller, CWE Inc., USA). The head was shaved and skin prepared with antibacterial soap, isopropyl alcohol, and betadine iodine solution. A 3 cm midline incision was made from the eyes, then the skull was cleared of periosteal tissue. A burr hole was created (Micromotor drill, Stoelting Co., USA) at AP −4 mm, ML 2.5 mm, for viral injection using a Neurosyringe 1701 (#65460-05, HamiltonCompany, USA). One microlitre of the virus was administered at two cortical depths over 15minutes, at DV −1000 um and at DV −500 um, for a total of 7.2 × 10⁹ vc. The incision was cleaned and sutured with 4-0 Polysorb (Covidien #SL-5637), and the animal then placed in a pre-warmed recovery cage.