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Journal: iScience
Article Title: Toxoplasma gondii -induced region-specific disintegration in glutamatergic neurotransmission is linked to cognitive impairments in mice
doi: 10.1016/j.isci.2025.114415
Figure Lengend Snippet: Indirect modulation of neuroinflammatory and immune cell responses through targeted parasite burden reduction by sulfadiazine in chronic T . gondii infection (A) Representative gating strategy for flow cytometric analysis of mouse brains chronically infected with T . gondii . Cells isolated from the whole brain tissue were first selected according to their size and granularity in the forward and side light scatterplots (FSC-A, SSC-A, not shown), and only live single cells were gated. Microglia were identified by CD11b + CD45 int expression, while myeloid cells were first gated on CD11b + CD45 high , then further subdivided into Ly6G + neutrophils and Ly6G − mononuclear cells. Mononuclear cells were further classified based on Ly6C expression. Lymphocytes were identified through CD11b − CD45 high expression and further distinguished by NK1.1 + and CD3 + markers, with CD3 + T cells categorized into CD4 + T helper and CD8 + cytotoxic subsets. (B) Bar charts show the absolute cell count of peripheral immune cells recruited to the brains of T . gondii and T . gondii +Sulfa groups. (C) Z score heatmap representing standardized median fluorescence intensity (MFI) values of microglial surface markers across experimental groups. Asterisks denote statistical significance based on raw MFI values, highlighting differential expression following T . gondii infection and sulfadiazine treatment. (D) Relative mRNA levels of phagocytosis-associated markers CD36 and TREM2, with fold changes normalized to Hprt and adjusted to the mean values of the T . gondii -infected group. (E) Relative mRNA expression levels of cytokines in the brains of infected and treated mice, normalized to Hprt . (F) Expression of T . gondii stage-specific markers in brains of chronically infected mice. SAG1 encodes the major surface antigen of tachyzoites, while BAG1 is specific to the bradyzoite stage, indicating establishment of chronic infection. Expression values are normalized to Gapdh . n = 5 per group. Statistical analysis: for plots (B, D, E, and F), comparisons between T . gondii -infected and sulfadiazine-treated groups were performed using the Mann-Whitney U test. For plot (C), data were analyzed by one-way ANOVA followed by Holm-Šidák post hoc test for predefined comparisons between T . gondii -infected and sulfadiazine-treated groups. Bars represent the mean values ±standard error of the mean (∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗ p < 0.01).
Article Snippet: TaqMan Gene Expression Assay Trem2 ,
Techniques: Infection, Isolation, Expressing, Cell Characterization, Fluorescence, Quantitative Proteomics, MANN-WHITNEY