For all tissue samples (except as stated below), used for molecular and bioenergetics evaluations, rats were placed in a plastic chamber connected to a vaporizer machine (Dispomed, Joliette, QC, Canada) to be first anesthetized with inhaled Isoflurane USP (Batch number: N0G120H20A; Fresenius Kabi, Toronto, ON, Canada; 4 vol% in O2; ratio 4:1).8 (link)–10 (link) Sedation was monitored through the absence of painful response to pedal and toe pinch. Rats were then rapidly killed by decapitation.
For electron microscopy studies, rats were anesthetized with the intraperitoneal injection of a mixture of ketamine (Vetalar, 90 mg/kg; Bioniche Animal Health Canada, Inc, Belleville, Ontario) and xylazine (Rompun, 10 mg/kg; Bayer Healthcare LCC, Whippany). Plane anesthesia was confirmed by the loss of pedal and toe pinch reflex. Animals were then placed in dorsal position near the peristaltic pump for transcardiac perfusion, which consists in an end-point procedure. All animals were killed following the guidelines of the Canadian Council on Animal Care and the US National Institutes of Health Guide for the Care and Use of Laboratory Animals to ensure minimum pain, suffering, and distress.
For electron microscopy studies, rats were anesthetized with the intraperitoneal injection of a mixture of ketamine (Vetalar, 90 mg/kg; Bioniche Animal Health Canada, Inc, Belleville, Ontario) and xylazine (Rompun, 10 mg/kg; Bayer Healthcare LCC, Whippany). Plane anesthesia was confirmed by the loss of pedal and toe pinch reflex. Animals were then placed in dorsal position near the peristaltic pump for transcardiac perfusion, which consists in an end-point procedure. All animals were killed following the guidelines of the Canadian Council on Animal Care and the US National Institutes of Health Guide for the Care and Use of Laboratory Animals to ensure minimum pain, suffering, and distress.