MORPHOGLIA: MAPPING OF GLIAL MORPHOLOGIES AND SPATIAL ORGANIZATION

Microglia and astrocytes are dynamic central nervous system cells whose functions are tightly linked to their morphology and spatial organization, particularly under neuroinflammatory conditions. However, existing analytical approaches often rely on predefined categories and fail to capture subtle, continuous variations in glial phenotypes and their spatial interactions.
Here, we present MorphoGlia, a pipeline for the unbiased characterization and spatial mapping of glial morphologies. MorphoGlia employs a data-driven approach to select morphometric features, perform dimensionality reduction, cluster morphological states, and map clustered cells back onto tissue architecture, providing integrated morpho-spatial context.
We first applied MorphoGlia to microglia in an Alzheimer’s model, comparing groups in the CA1 and Hilus hippocampal subregions. The pipeline reliably distinguished treatment-dependent microglial phenotypes and identified spatially localized pro-inflammatory morphology clusters enriched in Alzheimer’s animals.
We next extended this framework to astrocytes, integrating morphology-based clustering with spatial graph analysis and point-process statistics to investigate how astrocyte organization adapts to graded neuroinflammatory stress. Despite inflammation-induced increases in astrocyte density, regional morphological identities and characteristic interaction scales were preserved through coordinated mesoscale reorganization into modular spatial structures. Computational modeling further demonstrated that this hierarchical buffering mechanism emerges from fixed interaction rules and predicts collapse under severe pathology.
Together, these results establish MorphoGlia as a versatile platform for multiscale analysis of glial morphology and spatial organization. By bridging single-cell morphology with tissue-level topology, MorphoGlia enables systematic investigation of glial heterogeneity, adaptive reorganization, and failure modes in neuroinflammatory and neurodegenerative conditions.