Investigating Collagen IV Mutations in Mice

Young adult (3-mo-old) and middle-aged (12-mo-old) male and female littermate Col4a1^+/+ and Col4a1^+/G394V mice were used in this study. The Col4a1^G394V mutation was backcrossed to C57BL/6J mice for over 20 generations (13 (link), 42 (link)). Animals were maintained in individually ventilated cages (≤5 mice/cage) with ad libitum access to food and water in a room with controlled 12-h light and dark cycles. All animal care procedures and experimental protocols involving animals complied with the NIH Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committees at the University of Nevada, Reno, and UCSF. mice were euthanized by decapitation and exsanguination under isoflurane anesthesia (Baxter Healthcare). Brains were isolated and placed in ice-cold Ca²⁺-free physiological saline solution (Mg-PSS, containing 140 mM NaCl, 5 mM KCl, 2 mM MgCl₂, 10 mM HEPES, and 10 mM glucose (pH 7.4, NaOH), supplemented with 0.5% bovine serum albumin).