Generation of Knockout Mice Sample Clauses

Generation of Knockout Mice. (KO) and Maintenance of Mouse Colony Gpr37 knockout mice (Gpr37-/-) were obtained from Xxxxxxx Laboratory (strain Gpr37tm1Dgen, stock number 005806) and GPR37L1 knockout mice (Gpr37L1-/-) were obtained from the NIH Mutant Mouse Regional Resource Centers (strain Gpr37l1tm1Lex, stock number 011709-UCD). These mouse lines were backcrossed with wild-type C57BL/6J mice (Xxxxxxx Laboratory) for 10 generations each to ensure uniformity of genetic background. Following this backcrossing, Gpr37-/- and homozygous Gpr37L1-/- mice were crossed to generate the Double-KO (DKO) line of mice. Genetic deletion of Gpr37 and/or Gpr37L1 was confirmed by DNA sequencing, and loss of GPR37 and/or GPR37L1 protein expression was confirmed by Western blotting of brain tissue samples with specific anti-GPR37 and anti-GPR37L1 antibodies (MAb Technologies). All mice were maintained on a C57BL/6J background and housed on a 12-h light/dark cycle, with food and water available ad libitum. All experiments were conducted during the light cycle prior to 4:00 p.m. and were performed in accordance with the guidelines of the Institutional Animal Care and Use Committee of Emory University. Wild-type (WT) littermates were used as controls for all single mutant experiments, and age-matched WT littermates from each single mutant line were combined and used as controls for experiments involving DKO mice. Male mice were used in all seizure induction paradigms. In all experiments, the experimenter was blinded to the genotypes of the animals.