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. 2021 Jul 12;8(4):ENEURO.0559-20.2021. doi: 10.1523/ENEURO.0559-20.2021

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Copyright © 2021 Bordone et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Figure 4. — Experimental design to evaluate the reversion of LID by miRNA-Fyn treatment. A, Timeline of the post-L-DOPA treatment schema: 6-OHDA lesion into the MFB followed by the first L-DOPA trial for 15 d and AIMs analysis at indicated time points; striatal miRNA LV injection, and second L-DOPA trial, AIMs evaluation, and postmortem analyses. B, Cylinder test performed after 6-OHDA lesion, prior to random assignment of mice to treatment groups. Data are mean ± SEM. Kruskal–Wallis test H₍₂₎ = 3.612; p = 0.1643. C, Sum of values for axial dystonia, orolingual, and limb dyskinesia (total AIMs score) per day throughout the L-DOPA treatment. Experimental groups: non-injected (no LV; n = 9), injected with LV carrying a control (n = 5) or with miRNA-Fyn (n = 19). Data are mean ± SEM. Two-way ANOVA with repeated measures performed on data from the second L-DOPA trial (interaction: F_(8,120) = 2.520, p = 0.0144; time: F_(4,120) = 4.724, p = 0.0014; treatment: F_(2,30) = 1.089, p = 0.3495; subject: F_(30,120) = 38.32, p < 0.0001) and post hoc Tukey’s test did not show statistical differences between treatments. D, Percentage of fall after treatments calculated for each individual mouse. Second L-DOPA trial AIM scores compared with AIM score in the first L-DOPA trial for the same group of treated mice. Data are mean ± SEM. One-way ANOVA (F_(2,30) = 0.2390 p = 0.7889). E, Dot plot of data showing the distribution of the AIMs score within each group. The dotted line separates the first from the second L-DOPA trial. F, F’, Percentage of fall after treatments analyzed by estimation statistic and shown as a Cumming estimation plot. F, Raw data are presented as a swarmplot and mean ± SD are represented on the right of each experimental group. F’, Unpaired mean difference for two comparisons against the shared control (no LV group). The unpaired M_diff are plotted as bootstrap sampling distributions. Each mean difference is depicted as a dot. Each 95% CI is indicated by the ends of the vertical error bars. Unpaired M_diff (control LV vs no LV) = −5.99 and 95.0% CI [−16.0, 22.8] and unpaired M_diff (miRNA-Fyn vs no LV) = 7.89 and 95.0% CI [−9.89, 27.0] followed by two-sided permutation t test with p = 0.575 and p = 0.521, respectively.

Figure Contributions: Melina P. Bordone performed 6-OHDA lesions and Ana Damianich striatal lentiviral injections. Melina P. Bordone, Ana Damianich, Tomas Eidelman, and Juan E. Ferrario made animal nursing and cylinder tests. Melina P. Bordone, Tomas Eidelman, and Sara Sanz-Blasco scored LIDs. Melina P. Bordone performed statistical analyses and Melina P. Bordone, Juan E. Ferrario, and M. Elena Avale analyzed and discussed data.