Journal: Journal of Neuroinflammation

Article Title: TYROBP/DAP12 knockout in Huntington’s disease Q175 mice cell-autonomously decreases microglial expression of disease-associated genes and non-cell-autonomously mitigates astrogliosis and motor deterioration

doi: 10.1186/s12974-024-03052-4

Figure Lengend Snippet: Tyrobp deletion reduces C1Q and GFAP levels in Q175 mice. A RT-qPCR of C1q mRNA and western blot and densitometric analysis of C1Q protein in the striatum of WT and Q175 mice with and without Tyrobp (10 months of age), n = 4–6 mice per group. Quantitative analysis is shown as mean ± SEM. Each point represents data from an individual mouse. Statistical analysis was performed using Two-Way ANOVA. *p < 0.05; **p < 0.01. B Representative images showing GFAP (green) staining in the striatum of WT and Q175 mice with and without Tyrobp (upper panel). 4X magnification, Scale bar, 500 µm. Representative images (20X and inset, 40X) showing GFAP (green) staining in the striatum of WT and Q175 mice with and without Tyrobp . Quantification of the intensity is shown as mean ± SEM (n = 5–6 mice per group). Each point represents data from an individual mouse. Statistical analysis was performed using Two-Way ANOVA. *p < 0.05; **p < 0.01. Scale bar, 25 µm. C Heatmap showing Q175 vs WT (top) and Q175;Tyrobp(−/−) vs Q175 (middle) and Tyrobp(−/−) vs WT log 2 fold-change of reactive astrocyte marker genes. in the transcriptome nominal p-value from the differential expression analysis is shown. ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05

Article Snippet: C Heatmap of binomial p-values for Bioplanet pathways upregulated in human and mouse datasets.

Techniques: Quantitative RT-PCR, Western Blot, Staining, Marker, Quantitative Proteomics

Journal: Journal of Neuroinflammation

Article Title: TYROBP/DAP12 knockout in Huntington’s disease Q175 mice cell-autonomously decreases microglial expression of disease-associated genes and non-cell-autonomously mitigates astrogliosis and motor deterioration

doi: 10.1186/s12974-024-03052-4

Figure Lengend Snippet: Analysis of the HD microglia-specific transcriptome and the consequences of Tyrobp deletion. RNA was purified from freshly isolated striatal microglia and then sequenced. A Analysis of global levels of cell type-specific transcripts from sorted mouse microglia RNA-seq samples using geometric mean of the normalized counts of cell type marker genes of Drd1 (direct)-MSNs, Drd2 (indirect) -MSNs, astrocytes and microglia. B Volcano plot illustrating the DEGs identified in Q175 vs WT comparison. Only the genes with FDR < 0.1 are shown. C Heatmap of binomial p-values for Bioplanet pathways upregulated in human and mouse datasets. D Predicted upstream regulators of upregulated genes from Q175 microglia. E Intersection between genes with H3K27me3 marks from striatal microglia and upregulated genes from Q175 microglia. F Pathways associated with upregulated DEGs with a H3K27me3 mark. Color denotes database (Brown: Bioplanet; Green: KEGG pathways; Blue: Wikpathways). G Volcano plot illustrating the DEGs identified in Q175; Tyrobp (−/−) vs Q175 comparison. Only the genes with FDR < 0.1 are shown. H Heatmap of binomial p-values for Bioplanet pathways decreased in Q175; Tyrobp (−/−) , and integration with increased pathways from specified human and mouse datasets. I KEGG pathways enriched for upregulated genes in the microglial cluster of HD human snRNA-seq (pink/gray), integrated with the pathways downregulated by Tyrobp deletion in Q175 mice (blue). J Western blot and densitometric analysis of phosphorylated and total Erk protein in striatal samples. n = 5–6 mice per group. Quantitative analysis is shown as mean ± SEM. Each point represents data from an individual mouse. Statistical analysis was performed using Two-Way ANOVA. *p < 0.05

Article Snippet: C Heatmap of binomial p-values for Bioplanet pathways upregulated in human and mouse datasets.

Techniques: Purification, Isolation, RNA Sequencing, Marker, Comparison, Western Blot

Journal: Journal of Neuroinflammation

Article Title: TYROBP/DAP12 knockout in Huntington’s disease Q175 mice cell-autonomously decreases microglial expression of disease-associated genes and non-cell-autonomously mitigates astrogliosis and motor deterioration

doi: 10.1186/s12974-024-03052-4

Figure Lengend Snippet: Tyrobp deletion reduces pathways activated in human HD brain but not in HD mouse models. Bulk RNA-seq was performed on whole striatal RNA. A The number of DEGs for each data set at adjusted p < 0.1 and nominal p < 0.05 is shown. Blue bars indicate downregulated DEGs, and red bars indicate upregulated DEGs. B GO terms (biological process) associated with genes from Q175 vs WT data sets after GSEA analysis. C Cell-type enrichment analysis for Q175; Tyrobp (−/−) vs Q175 DEGs (nominal p-value < 0.05). Lines represent − log 10 p-value of chi-square calculation of the cell type enrichment. D GO terms (biological process) associated to genes from Q175; Tyrobp (−/−) vs Q175 data sets after GSEA analysis. dSNP = direct (Drd1) spiny neurons; iSNP = indirect (Drd2) spiny neurons ( E ) Heatmap of Normalized Enrichment Score for Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways detected in human and mouse datasets. Pathways have been ranked based on descending normalized enrichment score from the Human HD BA9 dataset (Agus et al., 2019 ). Column #1: Human HD Brodmann area 9 (BA9) (Agus et al., 2019 ); #2: Human HD Cingulate Cortex (Al-dalahmah et al., 2020 ); #3 Presymptomatic human HD BA9 vs control (Agus et al., 2019 ); #4: Human HD caudate (CAU) v Control (Elorza et al., 2021 ); #5 Presymptomatic Human Caudate (Agus et al., 2019 ); #6: Q175; Tyrobp (−/−) vs Q175; #7: Q175 vs WT. F GSEA results for Cytokine-Cytokine receptor interactions and Complement and coagulation cascade pathways. Normalized gene scores are shown

Article Snippet: C Heatmap of binomial p-values for Bioplanet pathways upregulated in human and mouse datasets.

Techniques: RNA Sequencing, Control, Coagulation