Single nucleus RNA-seq of subcortical white matter from lysophosphatidylcholine (LPC) and cuprizone toxicity mouse models

We compared the transcriptional changes in the cells of the corpus callosum, focusing on the oligodendrocyte and microglia, between LPC and cuprizone mediate demyelination. We further compared the transcriptional phenotypes to human MS patient samples. We identified distinct disease-associated oligodendrocyte states with shared pathological changes to MS, and observed a common but altered remyelination state between the models. Microglia response to demyelination is highly conserved, but human MS-associated microglia exhibit significantly more heterogeneity than we found in the mouse models. (doi: https://doi.org/10.1101/2025.03.24.645058) Overall design: LPC: Adult (8-12wk) C57BL6 mice were injected with 1% LPC into the corpus callosum of one brain hemisphere. Saline was injected on the contralateral side. 7 days later, mice were injected with Neutral Red dye by IP 2 hours prior to sacrifice. Mice were anesthetized, perfused with cold PBS for 5mins, and the brain was dissected out into cold PBS. Coronal slices of the brain were made and corpus callosum lesions were identified by the presence of Neutral red stain. The corpus callosum lesion was dissected out and the corresponding region on the contralateral side was also collected. Tissue was placed in Eppendorf tubes, excess liquid was removed as much as possible, and tissue was flash frozen in liquid nitrogen and stored at -80 until single nuclei processing. Cuprizone: Adult (8-12wk) C57BL6 mice were fed 0.2% cuprizone chow for 5 consecutive weeks. Control mice were fed normal chow. Mice were anesthetized, perfused with cold PBS for 5mins, and the brain was dissected out into cold PBS. Coronal slices of the brain were made. The corpus callosum was dissected out for cuprizone-fed mice and normal chow-fed mice. Tissue was frozen as above, and stored at -80 until processing.