Genetically Modified Mice in Stroke Research

All procedures fulfilled the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines on experimental design, animal allocation to different experimental groups, blinding of samples to data analysis, and reporting animal experiments. The GphnS268A/S270A mutant mouse was generated using CRISPR-Cas9 (Cyagen, USA) in BL6 background. These mutant mice develop normally and breed with Mendelian ratio. Heterozygous breedings were used to generate GphnS268A/S270A homozygous mutant mice; however, control C57Bl6/J-Crl1 mice were purchased from Charles River Laboratories (Germany). B6.129P2(C)-Cx3cr1tm2.1(cre/ERT2) (stock no. 020940) (57 (link)) mice and Bdnftm3Jae or BDNF^Tg (stock no. 004339) (58 (link)) were obtained from the Jackson Laboratory, and heterozygous breeding pairs were set up to generate BDNF^wt/wt/CX3CR1^ERT2Cre+/− and BDNF^flox/flox/CX3CR1^ERT2Cre+/-cKO lines. We compared results within same genotypes. The mice were injected (intraperitoneally) on four consecutive days with tamoxifen dissolved in corn oil (H-6278, Sigma-Aldrich; 1 mg/day) to induce Cre expression at 4 weeks, followed by sham or MCAO surgery at 8 to 9 weeks of age. Animals were randomly assigned, and both genders were used for both conditions. The PLX5622 treatment for microglia depletion followed the recommended company dose (1200 mg of active form of PLX5622/kg of chow).