Figure 1.

α-syn PFF-induced PD like pathology is rescued by NLY01. (a) Schematic diagram of the α-syn PFF experimental design. (b) Representative double-immunostaining for p-α-syn^ser129 (p-α-syn) (green) and TH (red) in the SNpc. White arrows point to DA neuron loss. Low power images were generated by the tile scan algorithm in the Zen software (n=6, biologically independent animals). Scale bar, 100 μm or 10 μm, respectively. (c) Percentage of TH neurons with p-α-syn^ser129 positive inclusion in the SNpc region. Error bars represent the mean ± S.E.M. (n=6, biologically independent animals, p value = 0.0007). Unpaired two-tailed Student’s test were used for statistical analysis. (d) Representative photomicrographs from coronal mesencephalon sections containing TH-positive neurons in the SNpc region (n=10, biologically independent animals). Scale bar, 500 μm. (e) Unbiased stereologic counts of TH and (f) Nissl-positive neurons in the SNpc region. Error bars represent the mean ± S.E.M. (n=10, biologically independent animals). (g) HPLC assessment of DA concentrations in the striatum of PBS and α-syn PFF stereotaxic injected mice that were treated with vehicle or NLY01. Error bars represent the mean ± S.E.M. (n=5, biologically independent animals). (h-i) Behavioral tests after PBS or α-syn PFF stereotaxic intrastriatal injection at six-months in vehicle or NLY01 treated mice. (h) Amphetamine rotation test, and (i) pole test. Error bars represent the mean ± S.E.M. (n=12 PBS with vehicle, n=13 PBS with NLY01, n=14 α-syn PFF with vehicle, and n=11 α-syn PFF with NLY01, biologically independent animals). Two-way ANOVA was used for statistical analysis followed by Tukey’s multiple comparisons test. ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 vs. PBS stereotaxic injected mice with vehicle; *P < 0.05, **P < 0.01, ***P < 0.001 vs. α-syn PFF stereotaxic injected mice with NLY01. Maximum time to climb down the pole was limited to 60 sec.