Antisedan

MI was induced as previously described.^{51 (link)} Mice were deeply anesthetized with an intraperitoneal injection of MMB (0.1 mL per 10 g). Overall, the mice were intubated using a small rodent ventilator (MiniVent 845, Harvard Apparatus, Holliston, MA). Left lateral thoracotomy was performed, and the LAD coronary artery was ligated using an 8/0 nylon suture after removing the pericardium. The chest was closed, antipamezole (0.75 mg·kg⁻¹; Antisedan; Nippon Zenyaku Kogyo Co., Ltd.) was administered for reversal, and then the mice were removed from the respirator. The mice were allowed to recover on a warm surface. Sham mice underwent all procedures except for actual LAD occlusion. Some mice were examined until 28 days after MI to analyze survival. The other mice were examined until 7 days after MI and then sacrificed to obtain samples. All mice were necropsied to obtain evidence regarding post-MI cardiac rupture or heart failure as described previously.^{52 (link)}

The maxillary molars were extracted under anesthesia with triple anesthesia (medetomidine hydrochloride 0.75 mg/kg (Domitol, Nippon Zenyaku Kogyo Co.,Ltd. Fukushima, Japan), midazolam 4.0 mg/kg (Dormicum, Sandoz K. K., Tokyo, Japan), butorphanol 5.0 mg/kg (Vetorphale, Meiji Seika Pharma Co., Ltd. Tokyo, Japan)) using hooked-end forceps. After surgery, mice were injected with Atipamezole 0.75 mg/kg (antisedan, Nippon Zenyaku Kogyo Co.,Ltd.) to antagonize the anesthesia effect. Two weeks later, the tissue from the extraction socket was collected and used for analysis.

Stab wound surgery was performed using a modified version of the procedure previously described by Hashimoto et al.^{18 (link)}. Mice (6 weeks old) were anaesthetised by a mixture of 0.75 mg/kg Domitor (Nippon Zenyaku Kogyo, Fukushima, Japan), 4 mg/kg midazolam (Sandoz, Holzkirchen, Germany), and 5 mg/kg Vetorphale (Meiji Seika Pharma, Tokyo, Japan). A 19-gauge needle was then used to penetrate the skull in the occipital region of the right hemisphere, and a 27-gauge needle was inserted through this hole along the rostrocaudal axis, and the needle was pulled out gently to induce just one stab wound to the right cerebral cortex. After the operation, the Domitor antagonist Antisedan (0.75 mg/kg; Nippon Zenyaku Kogyo) was injected intraperitoneally to facilitate recovery from anaesthesia. A sample size calculation was based on our previous study^{18 (link)}.

Animals were pre-anesthetized with an intramuscular injection of 0.04 mg/kg of medetomidine (Domitor; Nippon Zenyaku Kogyo, Koriyama, Japan), 0.40 mg/kg of midazolam (Dormicam 10 mg; Astellas Pharma, Tokyo, Japan), and 0.40 mg/kg of butorphanol (Vetorphale; Meiji Seika Pharma, Tokyo, Japan). They were also administered 15 mg/kg ampicillin (Viccillin; Meiji Seika Pharma, Co., Ltd.) and hydrated subcutaneously with 2 mL/head of fluid (KN No.1 injection; Otsuka Pharmaceutical, Tokyo, Japan). Thereafter, animals were anesthetized by inhalation with 1.0–3.0% isoflurane (Forane; Abbott Japan, Tokyo, Japan) via a ventilation mask. Anesthetization management was performed by spontaneous respiration during the operation, monitoring the heart rate and the arterial oxygen saturation. After oocyte collection or embryo transfer, 0.20 mg/kg atipamezole (Antisedan; Nippon Zenyaku Kogyo) was administered intramuscularly into the animals. For postoperative analgesia and infection control, 1.2 mg/kg ketprofen and 15 mg/kg ampicillin were administered once daily for 3 consecutive days following the operations.