Fig 2: Effect of CRS on neurotransmitter and microglia cells in MMTV-PyVT mice. Effect of CRS on neurotransmitter in serum was reversed by antidepressant Xiaoyao pill (A,B,C,D). Immunofluorescence detection of microglia marker IBA1 (E) (200× above, 400× below). Relative expression levels of M1 polarization markers (CD86, IL-1ß, iNOS) and M2 polarization markers (CD206, IL-10, Arg-1) in microglia cells after CRS treatment (F). *, P<0.05; **, P<0.01. CRS, chronic restraint stress; CD86, cluster of differentiation 86; IL-1ß, interleukin 1 beta; iNOS, inducible nitric oxide synthase; SEM, standard error of mean.

Fig 3: Role of GLP-1 and TRF on induced decrease in Aß, GFAP, and IBA1 expression. (A–C) Representative hippocampus and cortex images from three groups stained for Aß (MOAB-2), GFAP, and (IBA1), including a 4× field and a 20× field. Scale bars are 200 and 50 µm respectively. (D–F) The quantitative analysis of the positive areas of the three proteins in the hippocampal and cortical regions, respectively. Hippocampus: Averages of 4 fields per slice, n = 4 animals per group were quantified. Cortex: Averages of 5 fields per slice were quantified. n = 4. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Aß: The Kruskal-Wallis test. GFAP: The Kruskal-Wallis test was used for quantitative analysis of hippocampal regions and Brown-Forsythe and Welch ANOVA tests for cortical regions. IBA1 of hippocampus: One-Way ANOVA followed by Dunnett’s test. IBA1 of cortex: The Kruskal-Wallis test.

Fig 4: BXD32 mouse retinas have advanced glial hypertrophy and an increased presence of monocytes in the outer retina. WT and BXD32 retinal sections were immunolabeled for glial cells using IBA1 (red), GFAP (green), and GS (blue). Beginning at the earliest timepoint examined p63 (A), BXD32 retinas exhibit dramatic upregulation of GFAP and a larger number of monocytes, presumably microglia, both in the outer retina and the retina as a whole. By p105 (B), no decrease in GFAP is observed and monocyte numbers are even higher than at p63. At p168 (C), the outer retina is mostly degenerated and while monocyte numbers are lesser from p105, GFAP is still upregulated, indicating continued retinal stress. GS levels remained relatively similar amongst WT mice and BXD32 mice, with occasional variations with age. Nuclei stained with DAPI (white/grey). RPE, retinal pigment epithelium; ROS/IS, rod outer segments/inner segments; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer. Scale bars = 20 µm.

Fig 5: Quantification of fIHC. Using ImageJ, mean intensity (MI) per unit area in arbitrary units (a.u.) was assessed for fIHC labeling of GFAP, GS, and IBA1 (A); NF-?B, NLRP3, and TNFa (B); and STAT3, pSTAT3-Y705 and S727, and SOCS3 (C) in WT (black) and BXD32 (grey) mouse retinas with age. Error bars = ±SEM. *, p < 0.05; **, p < 0.005.