Sevoflurane exposure was performed according to a previously described protocol.
30 (link) Briefly, sevoflurane exposure and/or laparotomy was started at ZT0. Mice were exposed to 2% sevoflurane in 22% oxygen for 2 h in an anesthesia chamber; the concentration was monitored with a gas outlet. The heart rate, blood‐oxygen saturation, and rectal temperature were monitored. The mice breathed spontaneously, and sevoflurane was well tolerated, with all monitored variables in the physiological range.
Laparotomy was aseptically performed with a method previously used in mice.
31 (link) Mice were anesthetized for 2 h with 2% sevoflurane and intracutaneously injected with 0.2% ropivacaine along the planned incision line. A 2‐cm vertical incision was made in the middle of the abdomen, the gastrointestinal tract was exteriorized and vigorously rubbed for 30 s, and the organs (liver, spleen, kidneys, and bowel) were gently probed with cotton for 30 min. The intestines were then placed back into the peritoneal cavity, and the skin was sutured with surgical staples. EMLA cream (2.5% lidocaine and 2.5% prilocaine, AstraZeneca, Sweden) was applied to the incision wound at the end of surgery and then every 8 h for 2 days for surgical pain relief. Body temperature was maintained with a heating pad during anesthesia/surgery.
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