Collagenase-Induced Intracerebral Hemorrhage in Mice

Animal studies were reviewed and approved by the Committee on Animal Use for Research and Education at Augusta University, in compliance with NIH and USDA guidelines. ICH was induced in adult male CD-1 mice (Charles River) as previously reported (Sukumari-Ramesh et al., 2012a (link),b (link), 2016 (link); Bonsack et al., 2016 (link); Sukumari-Ramesh and Alleyne, 2016 (link)). Briefly, mice (n = 72) were anesthetized with ketamine and xylazine and prone-positioned on a stereotaxic head frame (Stoelting, WI, U.S.A.). The body temperature was maintained at 37 ± 0.5°C during the surgical procedure using a small animal temperature controller (David Kopf Instruments, USA) and a burr hole (0.5 mm) was made 2.2 mm lateral to bregma using a high-speed drill (Dremel, USA) without damaging the underlying dura. A Hamilton syringe (26-G) containing 0.04U of bacterial type IV collagenase (Sigma, St. Louis, MO) in 0.5 μL phosphate buffered saline (pH 7.4; PBS) was inserted with stereotaxic guidance 3.0 mm into the left striatum to induce spontaneous ICH (Bonsack et al., 2016 (link)). After removal of the needle, the burr hole was sealed with bone wax and the incision was stapled. Sham mice underwent the same surgical procedure, but only PBS (0.5 μL) was injected.