Sprague dawley strain

Pregnant rats (E17) of the Sprague-Dawley strain were purchased from Envigo. Embryos were surgically removed from the mothers and cortices dissected in Hanks Balanced salt solution (Gibco). The meninges were removed, and cells were dissociated using a papain dissociation system (Worthington) before being resuspended in Neurobasal medium A supplemented with antibiotic-antimycotic solution (Gibco), L-glutamine substitute (GlutaMAX™; Gibco) and factor B27 (Gibco). Cells were plated on poly-D-lysine coated glass coverslips at a density of 5 × 10⁵ cells/well, or on poly-D-lysine coated six-well cell culture plates at a density of 2 × 10⁶ cells/well, and incubated at 37 °C in 5% CO₂ with half media changes every 3 days. Cells were transduced with A53T a-syn or EV AAVs at 2 days post-isolation at a MOI of 3000. After 3 days, media containing AAV vectors were removed. Cells were transduced with Scramble1.3 or PDpep1.3 AAVs at 2- or 5-days post-isolation at a MOI of 3000. Cells were fixed with 4% paraformaldehyde (PFA) for immunofluorescence staining or lysed for immunoblotting at 8 days post-isolation.

Male albino rats from the Sprague-Dawley strain (Envigo, Indianapolis, IN, USA) weighing 200–250 g (initial weight)—were maintained in a constant-temperature (23 °C), with a fixed 12-h light/12-h dark cycle and fed for 16 weeks ad libitum either a standard chow diet (SC, 27.0%, 13.0%, and 60.0% of calories provided by protein, fat, and carbohydrates, respectively) a HF, sucrose-enriched (HFS) diet (20.0%, 60.0%, and 20.0% of calories provided by protein, fat, and carbohydrates [sucrose], respectively) or a KD (20%, 80% and 0% of calories provided by protein, fat and carbohydrates, respectively). The energy densities for the SC, HFS and ketogenic diets were 3.43, 5.24 and 6.14 kcal/g, respectively. The SC diet (standard rat chow, catalog # 5012) was purchased from TestDiet (Richmond, IN, USA). The HFS and ketogenic diets (catalog # D12492 and D03022101, respectively) were purchased from Research Diets Inc. (New Brunswick, NJ, USA). Energy intake was not significantly different between the three groups at weeks 0, 4 and 8 (Table 1). At week 16, the KD-fed animals had 17.9% and 18.1% lower energy intake than the SC and HFS-fed animals, respectively (Table 1).