H116 5mg

6-hydroxydopamine (6-OHDA; 6-hydroxydopamine hydrobromide with ascorbic acid: Sigma-Aldrich #H116-5MG) injections were made into the medial forebrain bundle of P120 female mice as previously described (Thiele et al., 2012 (link)). 30 min before 6-OHDA injection, a solution containing 0.5 mg/ml pargyline (Sigma Aldrich #P8013) and 2.5 mg/ml desipramine hydrochloride (Tocris #3067) was injected i.p. at a dose of 5 mg/kg pargyline and 25 mg/kg desipramine. 200 nl of freshly prepared 15 mg/ml 6-OHDA in sterile saline + 0.02% ascorbic acid were injected into the medial forebrain bundle (MFB, coordinates from bregma: M/L ± 1.2 mm, A/P 1.2 mm, D/V –4.90 mm). Adult female wild-type mice were used as younger mice and male mice showed poor recovery following injection. 250–350 µl of meloxicam (5–10 mg/kg dose) was injected subcutaneously as an analgesic.
Mice were monitored daily following the injection to ensure recovery. Kitten Milk Replacement (Santa Cruz #sc-362120) was fed to mice daily for up to 2 wk following the injection to aid recovery and meloxicam was injected subcutaneously to alleviate pain if necessary. Motor function was assessed using the cylinder test each week following the injection (see below). 4 wk following 6-OHDA injection, mice were quickly anesthetized using isoflurane and decapitated, and their brains were fresh-frozen as described above for FISH.

The PD rat model was established by injecting 6-hydroyx dopamine (6-OHDA) (H116-5MG, Sigma-Aldrich, MO, USA) into the one-sided medial forebrain bundle (MFB) target of the rat [19 (link)]. Dissolve 6-OHDA powder in 0.2% ascorbic acid to prepare 8 mg/ml 6-OHDA solution. Rats were anesthetized by intraperitoneal injection of 30 mg/kg of 3% sodium pentobarbital. After rats were anesthetized, their heads were shaved and fixed on the brain stereotaxic frame. Then, the rat head skin was sterilized with 75% alcohol, and the rat head skin was cut with scissors to find the location of the anterior fontanel, which was then used as the origin for positioning. The rats of the model group were injected with 25 μg 6-OHDA at MFB (A/P = 4.4, M/L = 1.2, and D/V = 7.8 mm). When using the cranial drill, avoid damaging the brain of rats due to excessive force, and the microsyringe was used to slowly insert to the brain. Before awakening, rats should use a thermal insulation pad to keep the rats' body temperature at 36.5°C. In order to prevent infection, penicillin (100,000 units) was injected intraperitoneally after the operation. The experimental operation in the sham group was exactly the same as that in the model group, except that 6-OHDA was replaced with an equal volume of saline solution. The rats in the control group were raised normally without any treatment.