Project description:To investigate cellular landscape of dural immune cells, dural immune cells from 30 P28 male mice and 30 P7 male mice were FACS sorted and single-cell RNA seqs were performed
Project description:To investigate heterogeneity of CNS border vasculatures, endothelial cells (EC) from 30 adult male mice were FACS sorted and single-cell RNA seqs were performed
Project description:To investigate heterogeneity of MHCII high CNS border-associated mononuclear phagocytes (BAMPs), MHCII high BAMPs from 30 adult male mice were FACS sorted and single-cell RNA seqs were performed
Project description:Deubiquitinating enzymes play crucial roles in various cellular activities, yet their involvement in central nervous system (CNS) vascularization and barrier function remains elusive. Canonical Wnt signaling is essential for proper CNS vascularization and barrier maintenance. Using a loss-of-function screening for Wnt signaling activity, we identified ubiquitin-specific peptidase 9 X-linked (USP9X) as a key regulator in brain endothelial cells (BECs). Endothelium-specific Usp9x knockout reduces Wnt-signaling activity, compromising CNS vascularization and barrier function during development. Activation of Wnt signaling rescues these defects. Mechanistically, we uncovered β-catenin as a direct substrate of USP9X, with USP9X catalyzing K48 polyubiquitin chains to stabilize β-catenin. In pathological models of impaired CNS vascular barrier function, including an intracerebral hemorrhage model and an oxygen-induced retinopathy model, loss of Usp9x intensifies barrier disruption, accentuating defects. This finding implicates USP9X as a critical regulator of CNS vascularization and barrier function through Wnt signaling, offering insights into CNS disease implications.
Project description:While macrophages in the meningeal compartments of the central nervous system (CNS) has been comprehensively characterized under steady state, studying their contribution to physiological and pathological processes has been severely hampered by the lack of specific targeting tools in vivo. Recent findings have shown that the dural sinus and its adjacent lymphatic vessels act as a neuroimmune interface. Notably, the cellular and functional heterogeneity of extrasinusoidal dural macrophages outside this immune hub is currently unclear. Therefore, we comprehensively characterized these cells using single-cell transcriptomics, fate mapping, confocal imaging, clonal analysis and transgenic mouse lines. Extrasinusoidal dural macrophages were clearly distinct from leptomeningeal and CNS parenchymal macrophages in terms of their origin, expansion kinetics and transcriptional profiles. Lastly, functional studies demonstrated that during autoimmune neuroinflammation, extrasinusoidal dural macrophages perform efferocytosis of granulocytes. Our results highlight a previously unappreciated myeloid cell diversity and provide insights into the brain’s innate immune system.
Project description:To investigate different responses of dural macrophage subsets to systemic viral infection, we collected the dura mater from mice 12 h after systemic LCMV infection and macrophages were sorted