SOD1 G93A experiments

Humans with SOD1 mutations have an approximately female/male ratio of 1 and because of this females were chosen for the SOD1^G93A model¹⁷.To test whether blocking CPT1 could modulate disease progression in ALS, we used the SOD1 G93A mouse model. B6.Cg-Tg(SOD1*G93A)1Gur/J mice were purchased from Jackson Laboratory (Bar Harbor, Main, USA)⁹⁰. Mice were kept in a high barrier facility (The Animal Facility, Aalborg University). B6.Cg-Tg(SOD1*G93A)1Gur/J male mice were breed with C57Bl/6J female mice. Genotypes were evaluated using DNA extracted from ear tissue punches. RT-qPCR was performed using previously described primer sequences according to protocol⁹¹. Transgenic SOD1^G93A female mice were used for experiments (n = 14). SOD1^G93A mice were randomized into treatment with either etomoxir (5 mg/kg dissolved in olive oil) or placebo (olive oil) daily by oral gavage from day 70 (baseline) (Fig. 4a). SOD1^G93A mice were evaluated with a neurological score system weekly as previously described⁹² and weighted two times a week. SOD1^G93A mice and ALS patients’ losses muscle strength as the disease progresses and therefore muscle strength were assessed weekly using a grip strength meter (Bioseb, France). Grip strength were measured in grams and normalized to weight according to protocol⁹³. Furthermore, neurological function and muscle strength were assessed using the hang-wire test as previously described^92,93. Briefly, mice were put on a wire lid and gently turn upside down. The time until the mice fell down was measured (latency to fall), cutoff time was set to 180 s. Each mice performed three trails and the highest value was used for statistics as previously described^92,94. Mice were euthanized when they reached a neurological score of 4 or above or if they had a weight loss of 20% or more according to ethical guidelines. The SOD1^G93A/Cpt1a mice (n = 6) were evaluated with the same procedures from day 70 until day 127 (Fig. 4f).