Total numbers of TH-positive and Nissl-positive neurons were estimated bilaterally from every 4th section through the extent of the SNpc of each brain. Stereology was performed at 400× using an Axioplan 2 imaging microscope (Zeiss, Göttingen, Germany) fitted with a DEI-750 CE video camera (Optronics, CA, United States) and a LEP MAC5000 motorized stage controller (Ludl Electronic Products, NY, United States). The software package used was Stereo Investigator (MBF Bioscience, VT, United States). The coefficient of error for the individual counts was 0.01. A counting frame of 50 μm × 50 μm and a height of 10 μm was chosen. Only the neurons within the counting frame were counted. At least 50 markers were counted within 50 framing sites for each mouse. The neuron count multiplied by four was used as the final number of TH+ cells. Data are expressed as TH(+)-dopaminergic neurons/SNpc (Healy-Stoffel et al., 2012; Park et al., 2012; Nam et al., 2015; Shen et al., 2017; Peng et al., 2018).

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