Mice were anesthetized with a combination of MMF (Medetomidin 0.5 mg/kg, Orion Pharma; Midazolam 5.0 mg/kg, Ratiopharm; Fentanyl 0.05 mg/kg, B.Braun). Mice were subjected to a laminectomy in order to expose the dorsal region of the spinal cord at cervical level 2 (C2). A unilateral lesion of the superficial dorsal column was made, using fine iridectomy scissors. Care was taken to avoid involving the underlying CST. Following surgery, an analgesic (Metacam, 1 mg/kg, Boehringer Ingelheim) was orally administered. Mice were kept on a heating pad at 38 °C until fully awake.
Medetomidin
Manufactured by Orion Pharma
Medetomidin is a laboratory equipment product used for the administration and monitoring of anesthesia in small animals. It is a potent alpha-2 adrenoceptor agonist that produces sedation, analgesia, and muscle relaxation. The product is intended for use by trained medical professionals in a controlled laboratory setting.
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5 protocols using medetomidin
1
Spinal Cord Injury Model in Mice
2
Surgical Procedures in Mice
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For all surgical procedures, mice were anesthetized with an i.p. injection of Midazolam/Medetomidin/Fentanyl (Medetomidin 0.5 mg/kg, Orion Pharma; Midazolam 5.0 mg/kg, Ratiopharm; Fentanyl 0.05 mg/kg, B.Braun) on a heating pad (38°C). For pain management, meloxicam (Metacam®, Boeringer Ingelheim) was administered 6 h after antagonization and every 12 h for 72 h.
3
Spinal Cord Hemisection in Mice
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Mice were anesthetized with MMF (medetomidin 0.5 mg/kg, Orion Pharma; midazolam 5.0 mg/kg, Ratiopharm; fentanyl 0.05 mg/kg, B. Braun). Once the mice presented no reflex reaction from paw pinching, their backs were shaved and a laminectomy was performed at T8 levels. The dura was exposed, and a dorsal hemisection was performed using irridectomy scissors38 ,58 (link) (Bradley et al.58 (link); Loy et al.59 (link)). After the hemisection, the wound was closed, and the skin was sutured. An antagonist mix was given (atipamezole 2.5 mg/kg, flumazenil 0.5 mg/kg, and naloxon 1.2 mg/kg), and mice were kept on a heating pad until completely awake. Mice received meloxicam (Metacam, 1.5 mg/ml oral suspension) at 12, 24, and 48 h following the injury.
4
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.
5
Controlled Cortical Impact Traumatic Brain Injury in Mice
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Mice were anaesthetized via an intraperitoneal (i.p.) injection of MMF (Medetomidin 0.5 mg/kg, Orion Pharma; Midazolam 5.0 mg/kg, Ratiopharm; Fentanyl 0.05 mg/kg, B.Braun). When pedal reflex disappeared, animals were fixed on the stereotactic frame (Precision Systems & Instrumentation, LLC). The skin was incised and a square craniotomy (4 mm × 4 mm), positioned between Lambda, Bregma, the sagittal suture and the side of the skull was made. The injury to the brain was induced using a TBI-0310 impactor (Precision Systems & Instrumentation, LLC), in the area of the somatosensory cortex (flat-edged rod of 3 mm diameter, velocity of 6 m/s, a dwell time of 150 m/s and a depth of 0.3–0.5 mm). The removed bone piece was re-positioned to its previous location and glued to the skull with Vetbond (3 M™ Vetbond™, 3 M United States). The skin was sutured. The mean apnea duration and righting reflex duration (time for a mouse to flip onto its feet from a supine position after the injury) were recorded in order to evaluate the severity of the injury.
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