B6 cg tg sod1 g93a 1gur j line

Male mice of the congenic B6.Cg-Tg(SOD1^*G93A)1Gur/J line, which was backcrossed for at least 10 generations to C57Bl/6J, were purchased from JAX (Stock No. 004435, The Jackson Laboratory, Bar Harbor, ME, USA). Male mice were bred in-house with C57Bl/6J females obtained from CRIVER (Charles River Laboratories, Sulzfeld, Germany). Mating generated hemizygous SOD1^G93A transgenic mice and non-transgenic littermate (ntg) controls. Progenies were genotyped for presence of the human SOD1 gene by a quantitative PCR assay of DNA obtained from ear markings, as previously described [71 (link)]. Copy numbers of the transgene were checked by calculation of the delta cycle threshold (∆CT) = CT_{internal control} − CT_{gene of interest}. Female SOD1^G93A mice with a high copy number of the transgene were selected for stratified randomisation into equal groups. As control, the genotype of each animal was confirmed by a second quantitative PCR at the end of the study. Housing of the animals was under the same terms at the animal facility of the Forschungszentrum Jülich as described previously [72 (link)]. Mice were housed in mixed-genotype in a controlled environment (12/12 h light/dark cycle, humidity maintained around 50% and a room temperature between 20 °C and 23 °C). Food and water were available ad libitum.

Male mice of the congenic B6.Cg-Tg(SOD1*G93A)1Gur/J line were purchased from JAX (Strain #: 004435, The Jackson Laboratory, Bar Harbor, ME, USA) and bred in-house with C57BL/6J females obtained from CRIVER (Strain Code 632, Charles River Laboratories, Sulzfeld, Germany). Each mouse was genotyped using RT-PCR with DNA extracted from ear tissue collected at P21. The SOD1 copy number was checked prior to the study start to ensure a stable phenotype progression as previously described [17 (link)].
The mice were housed at the animal facility of the Forschungszentrum Jülich under a controlled specific-pathogen-free (SPF) environment (12/12 h light/dark cycle, humidity maintained around 50%, and a room temperature between 20 and 23 °C). Food, water, and cages were autoclaved prior to entering the SPF area. A maximum of five SOD1*G93A mice and non-transgenic littermates were housed in individually ventilated cages on standardized rodent bedding (Rettenmaier, Rosenberg, Germany). Dried, pelleted standard rodent chow (Altromin, Lage, Germany) as well as tap water were available for the animals ad libitum. After disease onset, nutrition of the animals was ensured using gel pads (Solid Drink® SDST-75, Triple A trading, Tiel, The Netherlands).

All animal procedures were performed in accordance with the guidelines of the Australian National Health and Medical Research Council’s published Code of Practice for the use of animals in research and were approved by the Florey Institute of Neuroscience and Mental Health Animal Ethics Committee (permit number: 16-026). Transgenic HB9:GFP mice [B6.Cg-Tg(Hlxb9-GFP)1Tmj/J, stock no. 005029]^{19 (link)} and SOD1^G93A mice [B6.Cg-Tg(SOD1*G93A)1Gur/J line; stock no. 004435; RRID:IMSR_JAX:004435] were obtained from the Jackson Laboratory (Bar Harbor, ME, USA) and maintained on C57BL/6J backgrounds. SOD1^G93A male and HB9:GFP female mice were crossed to generate SOD1^G93A; HB9:GFP^+/− and HB9:GFP^+/− mice, which were subsequently crossed to generate experimental animals: SOD1^G93A; HB9:GFP^+/+ and HB9:GFP^+/+ mice. HB9:GFP^+/+ mice were henceforth called WT for this study. Mice were maintained under standard conditions of 12 h light/dark cycles with access to food and water ad libitum. Mice were time-mated overnight, and embryonic (E) Day 0.5 was designated following visualization of a vaginal plug the next morning.