Fig 1: Experimental timeline. The mice were administered Netrin-1 (45 μg/kg) or vehicle control (sterile PBS) at 1 h after surgery (Day 0). The mice in Cohort 1 were used to detect behavior 6 h after surgery. Once the behavior test was completed, they were sacrificed immediately for immunofluorescence and ELISA. The mice in Cohort 2 were used to detect behavior at 9 h after surgery. Once the behavior test was completed, they were sacrificed immediately for WB. The mice in Cohort 3 were used to detect behavior 24 h after surgery. Twenty mice in Cohort 4 were injected intravenously with 100 μl 10-kDa dextran at 6 h after surgery. Fifteen minutes after injection, each mouse was prepared for immunofluorescence. Another 20 mice in Cohort 4 were injected intravenously with 100 μl 10-kDa dextran at 24 h after surgery. Fifteen minutes after injection, each mouse was prepared for spectrophotometric quantification of 10-kDa dextran.
Fig 2: Exogenous recombinant human Netrin-1 decreased the levels of pro-inflammatory cytokines and increased the levels of anti-inflammatory cytokines in the sera, prefrontal cortex, and hippocampus of surgical mice. (A–C) Expression levels of IL-6, IL-10, and HMGB-1 in serum at 6 h after surgery/anesthesia. (D–F) ELISA was used to detect the expression of IL-6, IL-10, and Netrin-1 in the hippocampus at 6 h postoperatively. (G–I) Altered expression of IL-6, IL-10, and Netrin-1 in the prefrontal cortex at 6 h postoperatively. The data are presented as mean ± standard error of the mean for each group (n = 5 per cohort). *p < 0.05 vs. the control group, #p < 0.05 vs. the surgery group.
Fig 3: Earlier alterations of NTN‐1 and DCC in nigral dopaminergic neurons after MPTP intoxication. (A) Experimental design in a subacute MPTP mouse model. Brains were collected from mice at 6 h after 1 (MPTP‐1'd), 3 (MPTP‐3'd), or 5 (MPTP‐5'd) doses of MPTP injection and on day 7 (MPTP+7d) after the last injection. (B) Representative images of NTN‐1 (Green) in dopaminergic neurons (TH‐positive, Red) of the midbrain at 6 h after 1, 3, or 5 doses of MPTP injection and on day 7 after the last injection. Scale bar, 50 μm. (C) Representative images of DCC (Green) in dopaminergic neurons (TH‐positive, Red) of the midbrain at 6 h after 1, 3, or 5 doses of MPTP injection and on day 7 after the last injection. Scale bar, 50 μm. MPTP: 1‐Methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine; NTN‐1, netrin‐1; DCC: deleted in colorectal carcinoma; TH: tyrosine hydroxylase.
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