Transcriptomic profiling of oligodendrocytes, astrocytes, and microglia in a cuprizone-induced demyelination model

Demyelination is a central pathological feature of multiple sclerosis and many other neurodegenerative diseases, and the cuprizone diet is a widely used model for studying this process. This study examines the transcriptional changes in key glial cell types—oligodendrocyte and astrocytes—during demyelination induced by a 4-week cuprizone diet. Bulk RNA sequencing was performed on these cells isolated using magnetic-activated cell sorting (MACS) from mice on either a normal diet or a 0.3% cuprizone diet. The experimental groups included: (1) oligodendrocytes and astrocytes from mice on a normal diet, and (2) oligodendrocytes and astrocytes from mice on a cuprizone diet. This study provides a comprehensive transcriptomic view of how these glial cell types respond to demyelination and will enhance our understanding of their individual and collective roles in this pathological process. Overall design: Two-month-old C57BL/6 mice were fed either a standard chow diet (Naive) or a 0.3% cuprizone-enriched diet (CPZ) for 4 weeks to induce demyelination. Brain cell suspensions were prepared, and oligodendrocytes, astrocytes, and microglia were isolated using magnetic-activated cell sorting (MACS) with cell type-specific MicroBeads. Experimental groups: (1) Oligodendrocytes from mice on a normal diet (Naive_OL); (2) Oligodendrocytes from mice on a cuprizone diet (CPZ_Ctrl_OL); (3)Astrocytes from mice on a normal diet (Naive_Ast); (4)Astrocytes from mice on a cuprizone diet (CPZ_Ctrl_Ast). Bulk RNA sequencing was performed for all cell types and conditions. Each group has 3 or 4 repeats.