Overall, 3-month-old male Dnm1l+/- mice and their littermates were submitted to myocardial I/R in vivo. Dpr1+/- and WT mice were anesthetized by means of intraperitoneal sodium pentobarbital injections (80mg/Kg; Ceva Santé Animale) and received an injection of heparin (100IU/Kg, Heparine Choay®, Sanofi aventis, Paris, France) prior to thoracotomy. Myocardial I/R was achieved by temporarily occluding the left coronary artery, then releasing the occlusion as previously described (11,12). The duration of ischemia was 30 minutes, and that of reperfusion 24 hours.
To delimit the ischemic area-at-risk (AAR) and area of necrosis (AN), Evans Blue 4% (Sigma-Aldrich Co®, Missouri, USA) was injected through the left-ventricular apex. Thereafter, the heart sections were incubated with 2,3,5-triphenyltetrazolium chloride (TTC) (Sigma-Aldrich Co®, Missouri, USA) and fixed in 4% paraformaldehyde for 24 hours, as previously described [6 (link)]. The AN (white), AAR (not blue), and total left-ventricular (LV) areas from both sides of each section were measured using the software ImageJ 1.47. AAR/LV and AN/AAR were expressed as percentages, as previously reported [6 (link)].
To delimit the ischemic area-at-risk (AAR) and area of necrosis (AN), Evans Blue 4% (Sigma-Aldrich Co®, Missouri, USA) was injected through the left-ventricular apex. Thereafter, the heart sections were incubated with 2,3,5-triphenyltetrazolium chloride (TTC) (Sigma-Aldrich Co®, Missouri, USA) and fixed in 4% paraformaldehyde for 24 hours, as previously described [6 (link)]. The AN (white), AAR (not blue), and total left-ventricular (LV) areas from both sides of each section were measured using the software ImageJ 1.47. AAR/LV and AN/AAR were expressed as percentages, as previously reported [6 (link)].
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