Figure 3.

Single astrocyte co-localization with PSD-95 and analysis 24 h (PND 46) and 26-days (PND 72) after AIE. (A) Representative images of PSD-95 colocalized within 0.5 µm of a Lck-GFP⁺ astrocyte in the ACC, mPFC (IL and PL), LO-OFC, and VO-OFC 24 h after AIE. Scale bar 20 µm. (B) Quantification of puncta/astrocyte volume (puncta/µm³) in each of the PFC subregions. When comparing puncta number across subregions within the control groups, there was a significant increase in puncta number/astrocyte volume in LO-OFC and VO-OFC when compared to ACC and mPFC. There was no significant treatment effect on puncta number (H₂O vs. EtOH) within subregions. (C) Representative images of PSD-95 colocalized within 0.5 µm of an AAV+ astrocyte in the ACC, mPFC (IL and PL), LO-OFC, and VO-OFC 26-days after AIE. Scale bar 20 µm. (D). When comparing puncta number across subregions within the control groups, there was a significant increase in puncta number/astrocyte volume in LO-OFC and VO-OFC when compared to ACC and mPFC. There was a significant treatment effect on puncta number (H₂O vs. EtOH) demonstrated by a decrease in astrocyte-synapse co-localization in the ACC (p < 0.006) and the VO-OFC (p < 0.001). Data presented as box and whisker plots with interquartile range with mean, minimum, and maximum measures indicated along with individual data points. Analysis: two-way ANOVA with Tukey’s post hoc comparison, n = 5/treatment group.