PT  - JOURNAL ARTICLE
AU  - Ebert, Sarah N.
AU  - Eisner, Christine
AU  - Karamboulas, Konstantina
AU  - Bernier, Louis-Philippe
AU  - Kaplan, David R.
AU  - MacVicar, Brian A.
AU  - Miller, Freda D.
TI  - Single cell approaches define the murine leptomeninges:cortical brain interface as a distinct cellular neighborhood comprised of neural and nonneural cell types
AID  - 10.1523/ENEURO.0046-25.2025
DP  - 2025 Aug 08
TA  - eneuro
PG  - ENEURO.0046-25.2025
4099  - http://www.eneuro.org/content/early/2025/08/07/ENEURO.0046-25.2025.short
4100  - http://www.eneuro.org/content/early/2025/08/07/ENEURO.0046-25.2025.full
AB  - The interface barrier between the brain surface and the adjacent meninges is important for regulating exchanges of fluid, protein and immune cells between the CNS and periphery. However, the cell types that form this important interface are not yet fully defined. To address this limitation, we used single cell RNA-sequencing (scRNA-seq) and single cell spatial transcriptomics together with morphological lineage tracing and immunostaining to describe the cell types forming the interface barrier of the adult murine cortex. We show that the cortical interface is comprised of three major cell types, leptomeningeal cells, border astrocytes and tissue-resident macrophages. On the non-parenchymal side the interface is comprised of transcriptionally distinct PDGFRα-positive leptomeningeal cells that are intermingled with macrophages. This leptomeningeal layer is lined by a population of transcriptionally distinct border astrocytes. The interface neighborhood is rich in growth factor mRNAs, including many leptomeningeal ligands predicted to act on both the border astrocytes and macrophages. On the CNS side of the interface is the relatively cell-sparse cortical layer one containing interneurons, microglia, parenchymal astrocytes, oligodendrocyte precursor cells and oligodendrocytes. Except for the border astrocytes, layer one cells are not closely associated with the interface, suggesting that secreted ligands may be the major way the brain interface communicates with the underlying cortical parenchyma. Thus, our data provide a molecular/cellular resource describing the brain interface cell types and their interactions, thereby enabling future studies investigating how this distinct cellular compartment regulates CNS:periphery interactions.Significance Statement Recent years have seen significant progress in identifying the diverse cell types within the meningeal space. However, the mechanisms by which these cells interact with glial and neuronal cells in layer one of the adult murine cortex remain poorly understood. During development, communication between radial precursors and meningeal layers is crucial for proper brain formation, but the role of this interaction in adulthood is still unclear. Additionally, how resident immune cells in the leptomeningeal space signal to layer one cortical cells or meningeal mesenchymal cells during homeostasis remains an open question. Understanding the identity, location, and interactions of these cells is essential for unraveling the complex dynamics at this critical brain interface.