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2 protocols using naloxon

1

Surgical Anesthesia and Analgesia in Mice

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Mice were orally administered meloxicam (Metacam, 1.5 mg/ml, Boehringer Ingelheim) and anesthetized with intraperitoneal injections of MMF (medetomidin 0.5 mg/kg, Orion Pharma; midazolam 5.0 mg/kg, Ratiopharm; fentanyl 0.05 mg/kg, B. Braun) before surgical procedures. The depth of anesthesia was regularly controlled to ensure there was no reflex retraction from paw pinching. After surgery, mice were injected subcutaneously with an antagonist mixture (atipamezole 2.5 mg/kg, Providet Pharmaceuticals; flumazenil 0.5 mg/kg, Hameln Pharma GmbH; and naloxon 1.2 mg/kg, B. Braun). Prior to and after surgical procedures, mice were kept on a heating pad. Mice received meloxicam (Metacam, 1.5 mg/ml) orally for at least 3 d after the surgery.
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2

Viral Injections in Mouse Hippocampus

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Viral injections were performed under aseptic conditions in Prox1::cre or C57BL/6 mice up to 10 months old. The mice were anesthetized with a mixture of Fentanyl (Rotexmedica, Germany), Midazolam (Rotexmedica, Germany), and Domitor (Orion Pharma, Finland) via intraperitoneal injection ( 0.05/5.0/0.5  mg/kg ). Analgesia ( 0.05  mg/kg of buprenorphine; Buprenovet, Bayer, Germany) was administered intraperitoneally prior to the injection, and Xylocain (AstraZeneca, Germany) was used for local anesthesia. Stereotaxic injections were performed using an injection frame (WPI Benchmark/Kopf) and a microprocessor-controlled minipump (World Precision Instruments, Sarasota, Florida), 1000 nl of the viral solution was injected bilaterally into the hippocampus (rAAV-DIO-eGFP; rAAV-syn-GFP; coordinates: rostrocaudal: 2.1  mm from the Bregma; mediolateral: ±1.2  mm from the midline; dorsoventral: 2.1  mm ). After the injection, the scalp was sutured with PERMA-HAND Silk Suture (Ethicon), and an antibacterial ointment (Refobacin®, Almirall, Germany) was applied, followed by the intraperitoneal injection of a mixture of Naloxon (B. Braun, Germany), Flumazenil (B. Braun, Germany), and Antisedan (Orion Pharma, Finland) ( 1.2/0.5/2.5  mg/kg ). To prevent the wound pain, analgesia was administered on the 3 following days.
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