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. 2020 Oct 2;9:e58997. doi: 10.7554/eLife.58997

Figure 2. LRRK2 RC mutation restructures the nanoscale synaptic organization of dSPNs.

(A) Schematic depicting experimental design. (B) Structured Illumination super-resolution microscopy (SIM) image of dendritic spines on +/+, +/RC, and +/GS Drd1-Tomato expressing SPNs, labeled with antibodies to GluA1 (purple), and PSD95 (green). Open arrowheads, GluA1 nanodomains; arrowheads, PSD95 nanodomains. (C) Schematic diagram and object masks depicting GluA1, PSD95, and overlap nanodomains within a dendritic spine. Minimum distance between GluA1-PSD95 is measured from the closest edge of the two nanodomains, as shown. (D, E) Summary graphs and cumulative distribution of the minimum distance between GluA1 and PSD95 nanodomains. (F, G) Summary data and cumulative frequency for the overlap area of GluA1 and PSD95 nanodomains within dendritic spines. Asterisk in D and F reflect statistical significance for Tukey’s multiple comparison tests after one-way ANOVA, whereas asterisks in E and G show statistical significance for Bonferroni post-hoc comparisons after Kolmogorov-Smirnov tests. (H) Bar graphs showing the ratio of GluA1-PSD95 overlap area in E relative to PSD95 area, across genotypes. Data are represented as mean ± SEM. (I-K) Correlation plots of overlap in GluA1-PSD95 area versus PSD95 area for +/+, +/RC, and +/GS Drd1-Tomato expressing SPNs. *p<0.05, **p<0.01, ***p<0.001.

Figure 2—source data 1. Raw data plotted in graphs in Figure 2.

Figure 2.

Figure 2—figure supplement 1. LRRK2 mutations alter postsynaptic density area of SPNs.

Figure 2—figure supplement 1.

(A) Summary graph, PSD95 area across genotypes of dSPNs. Error bars reflect SEM. (B) Cumulative percentage of PSD95 area per dendritic spine across dSPN genotypes. (C) Summary graph, PSD95 area across genotypes of iSPNs. Error bars reflect SEM; asterisks, Tukey post-hoc comparisons after one-way ANOVA, +/+ vs. +/GS p<0.001 (D) Cumulative percentage of PSD95 areas per dendritic spine across iSPN genotypes.
Figure 2—figure supplement 1—source data 1. Numerical data of the graphs in Figure 2—figure supplement 1.
Figure 2—video 1. 3D Reconstruction of dendritic spine of an RC dSPN.
Download video file (5MB, mp4)
GluA1 (purple), and PSD95 (green), overlap area (white), as in Figure 2B. A surface mask is added for a fraction of the video, enabling the visualization of GluA1 and PSD95 in spine head.