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MB Sample ID: SA209249
| Local Sample ID: | Hgd83.1 + Nitisinone POS |
| Subject ID: | SU002265 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
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Collection:
| Collection ID: | CO002258 |
| Collection Summary: | A murine model of AKU was used for all experiments as described previously (Preston et al., 2014). Seventeen BALB/c Hgd-/- mice (Figure 5 for age and gender) were included in this study, 8 mice were administered nitisinone (4 mg/L) through their drinking water for 6 days, the remaining 9 mice received no nitisinone. All animals were housed in air conditioned rooms (with a 12 h dark/light cycle) at 20 °C and 53 % humidity, with access to food and water ad libitum at Liverpool John Moores University. All animal experiments complied with the ARRIVE guidelines and were carried out in accordance with the U.K. Animals (Scientific Procedures) Act, 1986 and associated guidelines, EU Directive 2010/63/EU for animal experiments. All mice were culled by carbon dioxide asphyxiation and CSF was removed from the cisterna magna following the puncture technique of Liu et al. (56), with some modifications. The cisterna magna was exposed by dissecting skin and overlaying muscles. Cauterisation was used to dry up any bleeding from surrounding tissues. A pulled glass pipette with an internal diameter of ~0.4 mm was attached to silicone tubing and connected to a 1 mL syringe with a 19 g needle. The tip of the glass pipette was used to puncture the membrane and held still just below the membrane by 1 person. Through a double-headed microscope a second person then used the 1 mL syringe to apply gentle pressure to encourage the CSF to flow up the capillary tube. Only clear CSF samples (i.e. non blood stained) were collected. Once collected CSF samples were transferred to clear Eppendorf tubes and stored at -80 °C until analysis. Samples were not acidified. The same glass pipette was used to collect CSF from each animal, and was washed with pure water between each animal. |
| Sample Type: | CSF |
