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, LTD) according to the user introduction. Rotary test was conducted based on the method of Vijitruth et al. [15]: mice were placed in a 20?cm diameter rotating bar, and the number of the rotation before mice left the bar was recorded. Each mouse was tested for five times at each time point and the interval between each time was 5?min. 2.3. The Effect of Low-Frequency rTMS on the Cortex Excitability of PD Model Mice Thirty-two eight-week-old male C57BL/6j mice weighing 240�C260?g were randomly divided into four experimental groups as described above, 8 mice for each group. Treatments were performed as described above. Then mice were assessed for their resting motor threshold (RMT) according to previous studies [16, 17] before agents injection and at 1st, 3rd, 7th, and 14th day after the last injection: mice were anesthetized with 40?mg/kg chloralhydrate and stimulated using selleck chemical Magpro X100 Magnetic Stimulator MS-275 datasheet (Dantec Dynamics A/S, Denmark) by placing coil at left brain of the mice. RMT values of the hind leg gastrocnemius were recorded by collecting the magnetic stimulation signal using Counterpoint Electromyography (Oxford Instruments, UK). 2.4. Effect of Low-Frequency rTMS on the Level of DA and Its Metabolites in the Striatum of PD Model Mice Twenty-four male C57BL/6J mice weighing 240�C260?g were randomly divided into four experimental groups as described above, 6 mice for each group. Treatments were performed on each group as described above. 24?h after the last stimulation of low-frequency rTMS, mice in each group were executed by breaking the necks, and the level of DA, homovanillic acid (HVA), and 3,4-dihydroxyphenylacetic acid (DOPAC) in mice striatum was detected using high performance liquid chromatography-electrochemical detection (HPLC-ECD) following standard procedures in Beijing Institute of Neurosciences, Capital Medical University. 2.5. Effect of Low-Frequency rTMS on the Function of NDN of PD Model Mice Fifty-six male C57BL/6J mice weighing 240�C260?g were randomly divided into four experimental groups as described above, 14 mice for each group. Treatments were performed on each group as described above. 24?h after the last stimulation of low-frequency rTMS, eight mice Quinapyramine from each group were anesthetized with 40?mg/kg chloralhydrate, perfused with 30?mL normal saline and 100?mL 40?g/L paraformaldehyde solution, and brain samples containing SN were then quickly frozen and cut into sections for immunohistochemical staining of tyrosine hydroxylase (TH), BDNF, and GDNF proteins in the SN: samples from each group were fixed with 3% hydrogen peroxide and blocked with 10% normal goat serum for 60?min. They were then incubated separately with primary rabbit antibodies against TH (1?:?5000, Chemicon), BDNF (1?:?100 ABcam), and GDNF (1?:?15, ABcam) at 4��C overnight. Samples were rewarmed for 60?min at 37��C before incubating with goat anti-rabbit IgG (1?:?100, Neomarker) for another 60?min at 37��C.
