Spatially Stereotyped Microgliosis Tracks Synaptic Pathology in the Demyelinated Superior Colliculus

Visual impairment is one of the most common and clinically salient manifestations of Multiple Sclerosis (MS), yet pathology across visual system structures remains incompletely defined. Although MS pathology has been extensively studied in the optic nerve, lateral geniculate nucleus, and visual cortex, involvement of the superior colliculus (SC), a key hub for visual processing, has not been systematically investigated. Here, we combined human postmortem tissue analysis with functional assessment and spatial mapping in the MS-relevant cuprizone (CPZ) mouse model to define how demyelination and secondary injury are organized within the SC. Postmortem SC tissue from donors with MS revealed myelin loss, including focal demyelinated lesions. In mice, CPZ treatment impaired visual function and induced widespread demyelination across SC layers, without detectable neuronal cell loss or axonal degeneration. Although diffuse demyelination was accompanied by widespread microgliosis characteristic of CPZ, atlas-based mapping uncovered a previously unrecognized spatial organization: a discrete high-microgliosis compartment that emerged in every CPZ-treated SC with strikingly stereotyped location and shape. This compartment did not correspond to canonical SC maps and was not explained by baseline differences in microglia or myelin or by variability in demyelination severity following CPZ. Instead, regions with elevated microgliosis showed a marked increase in synaptic elimination, suggesting that secondary synaptic pathology may contribute to the spatial organization of microgliosis beyond diffuse myelin loss alone. Prolonged CPZ exposure expanded the compartment in a stereotyped pattern, whereas CPZ withdrawal produced spatially ordered partial resolution while leaving a persistent high-microgliosis core concurrent with partial visual recovery. Together, these findings identify the SC as an MS-relevant site of injury and establish the CPZ-treated SC as a reproducible in vivo model for studying spatially patterned microglial reactivity, synaptic pathology, and incomplete inflammatory resolution after demyelinating injury.