Fvb mice

The experiments were performed in 12‐week‐old male wild‐type (FVB) mice purchased from Charles River Laboratories (Lecco, Italy). The mice were kept under constant temperature and humidity in a 12‐h controlled dark⁄light cycle. All the experiments were conducted within the animal welfare regulations and guidelines of the Italian national law D.L. 04/03/2014, n.26, about the use of animals for research.

Stereotactic injections with 1-μl AAV5-hSyn1-eNpHR3.0-YFP vector (5.14 × 10¹² vg/ml) were made in female FVB mice (Charles River, four to five weeks of age) housed before and after surgery in standard cages with 12/12 h light/dark cycle s and provided with food and water ad libitum. Animals were anesthetized with 1.5–2.5% isoflurane, 0.5 ml of bupivacaine was used as local anesthetic and chlorhexidine was used for wound cleaning. After placement in a stereotactic frame, a small skin incision and bore hole was made above the target region. A pulled glass capillary fitted to a 5-μl Hamilton syringe was lowered to the target region (anteroposterior (AP), −3.2 mm; mediolateral (ML), 3.1 mm from bregma; dorsoventral (DV), −3.6 and −3.2 from dura; 0.5 μl each depth, 0.1 μl/min). The wound was cleaned and closed with tissue glue. All procedures were performed in accordance with standard guidelines on experimental animal welfare and approved by the Malmö/Lund Animal Research Ethics Board (protocol number; MK 47-2015).

Animals rd1 mice were the albino FVB strain, which carries the Pde6b^rd1 allele (MGI: 1856373). BALB/c, CD1, and FVB mice were purchased from Charles River Laboratories. Due to availability, some of the FVB mice were purchased from Taconic, and we did not notice any difference between the two sources in terms of cone degeneration rate. C57BL/6J, rd10, and Parp1^-/- mice were purchased from The Jackson Laboratories and bred in house. C57BL/6J and rd10, which is on C57BL/6J background, carry a null mutation in the Nnt gene (Freeman et al., 2006 (link)), but not in the other strains (i.e., rd1, Rho^-/-, Parp1^-/-, CD1, and BALB/c). We crossed the Parp1^-/- mice with FVB mice to generate homozygous Parp1^-/-rd1 and Parp1^+/+rd1 mice. Genotyping of these mice was done by Transnetyx (Cordova, TN). The Rho^-/- mice were kindly provided by Janice Lem (Tufts University, MA) (Lem et al., 1999 (link)).

MuriGenics, Inc. (Vallejo, CA, USA) performed a thiamine–trimethoprim interaction study using 9- to 11-week-old (~20–25 g) FVB mice (n = 11) from Charles River Laboratories (Wilmington, MA, USA). Prior to the intervention, mice were fed a low thiamine diet (5 mg/kg) for one week followed by a thiamine-deficient diet (0 mg/kg) for the second week. The mice were then fasted for 16 h before treatment. On the day of the study, the mice were orally gavaged with either 2 mg/kg thiamine (n = 6; 3 female, 3 male) or a combination of 2 mg/kg thiamine and 61.5 mg/kg trimethoprim (n = 5; 3 female, 2 male). The brains were collected 3 h post intervention for thiamine vitamer analysis.