Fig. 5: Astrocytic exosomes increase neuronal miR-223 levels and downregulate miR-223-related neuronal gene expression. | Neuropsychopharmacology

Fig. 5: Astrocytic exosomes increase neuronal miR-223 levels and downregulate miR-223-related neuronal gene expression.

From: Exosomal secretion of a psychosis-altered miRNA that regulates glutamate receptor expression is affected by antipsychotics

Fig. 5

a Schematic of experimental design. b Graph showing mean ± SEM mature miR-223, miR-132, and miR-155 expression (based on mature miRNA qRT-PCR and normalized to highly expressed and unaltered let-7e—see Fig. S5a) in rat neurons treated with mouse astrocytic exosomes for 1–2 days relative to vehicle-treated rat neurons. c Graph showing mean ± SEM rat neuronal gene levels following mouse exosomal treatment for rat Grin2b, Gria2, and Adar2 mRNAs (all based on qRT-PCR and normalized to 18S rRNA—see Fig. S5b). Data are shown as ratios relative to the mean of vehicle. d Correlation between relative-to-vehicle miR-223 (mouse primer) and rat Grin2b mRNA expression. Spearman’s correlation coefficients and two-tailed p-values are shown in the graphs. Vehicle = purple circles, exosome treatment = green circles. e Graph showing mean ± SEM mouse mature miR-223 expression (based on mature miRNA qRT-PCR and normalized to let-7e) in mouse cortical neurons treated with mouse astrocytic exosomes for 2 days relative to vehicle-treated rat neurons. For (b, c, e) *p < 0.05, **p < 0.01, ***p < 0.001, based on two-tailed one-sample t test compared with vehicle. f Representative images following immunostaining of mouse cortical neurons with anti-Grin2b (red) and anti-MAP2 antibodies (green) after 2 days of treatment with astrocytic exosomes (Exosome) or no treatment (Vehicle). Scale bar = 50 µm. g Graph showing mean ± SEM relative to no-treatment vehicle Grin2b immunostaining intensity in mouse cortical neurons positive for MAP2 treated with mouse astrocytic exosomes for 1 day relative to vehicle-treated rat neurons (data from 3 to 4 images were averaged for each well/coverslip). **p < 0.01, based on two-tailed one-sample t test compared with vehicle. h, i Graphs showing mean ± SEM mouse neuronal gene levels following for mouse Grin2b (h) and Gria2 (i) mRNAs (all based on qRT-PCR and normalized to 18S rRNA) following 1 day of treatment with astrocytic exosomes treated with locked nucleic acid (LNA)-based miRNA inhibitors against mouse miR-223 (Anti-miR-223) or scrambled contol (Anti-Control)  astrocytic exosomes. j Graph showing mean ± SEM relative to no-treatment vehicle mouse Grin2b mRNA levels (based on qRT-PCR and normalized to 18S rRNA) following 2 days of treatment with exosomes from astrocytes treated with olanzapine or vehicle, or no exosome treatment. For (h–j): #0.05 < p < 0.10, *p < 0.05, **p < 0.01, ***p < 0.001, based on ANOVA with Tukey’s multiple comparisons test. The number of biological replicates is shown in each graph.

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