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. 2019 May 9;16(3):761–783. doi: 10.1007/s13311-019-00730-7

Fig. 8.

Fig. 8

Effect of CAL deficiency on rotenone-induced toxicity in vivo. (A) Following 4 weeks of AAV delivery, the SN sections were subjected to TH labeling (red) and the images with GFP immunofluorescence were obtained by confocal microscopy (up). Scale bar = 100 μm. The interference efficiency of AAV-shCAL was investigated by Western blotting (down, left) and quantification analysis was represented as the fold difference of the AAV-scramble group (down, right). (B) After virus injection, the animals underwent the apomorphine-induced rotational test to assess behavioral asymmetry. The total number of hemilateral rotational turns within 30 min was evaluated (up) (n = 7/ group) and the body weight during surgery was monitored (down) (n = 7/group). (C) TH-positive immunoreactivity was assessed by immunohistochemistry in the SN and striatum. Scale bar = 500 μm (SN), 1 mm (striatum). Quantification of TH-IR neurons in the SN or TH-IR fibers in the striatum represented the ratio of lesion side to intact side (n = 3 or 4/group). (D) The alterations in mGluR5, CAL, and p-JNK in the left (L) and right (R) striatum were assessed by Western blotting (up). Protein levels were normalized to GAPDH (TH, mGluR5, CAL) or total JNK (p-JNK) and represented as the ratio of lesion side to intact side by the expression of fold difference of the control group (n = 3/group). (E) mGluR5- and CAL-positive immunoreactivity in the SN was analyzed by immunohistochemistry (left) and the number of positive cells was analyzed (right). The quantification represented the percentage in the control group (n = 3 or 4/group). The black arrows represent positive cells that were stained brown (scale bar = 50 μm). Vehicle groups with AAV-scramble virus delivery were used as controls. Data shown in all panels of this figure represent the mean ± SD. Statistical significance was determined using Student’s t test (A) or 1-way ANOVA followed by Dunnett’s test (B–E). *p < 0.05, **p < 0.01, ***p < 0.001 versus control, #p < 0.05, ##p < 0.01, ###p < 0.001 compared to the rotenone lesion with AAV-scramble delivery group