Erythropoietin (EPO), a potent growth factor, named after its role in erythropoiesis, has been shown to not only be produced in the brain, but also to have significant effects in improving cognitive performance. The mechanisms by which EPO acts are yet to be fully explored, including the transcriptome changes elicited by it in the oligodendrocyte lineage. Here we explore these changes after a 3-week EPO/Placebo treatment using single-nuclei RNA sequencing. A total of 12.043 nuclei were classified into 5 cell types (oligodendrocyte precursor cell – OPC, committed oligodendrocyte precursor – COP, myelin-forming oligodendrocytes – MFOL, and 2 mature oligodendrocyte populations – MOL1 and MOL2), with a total of 637 unique differentially expressed genes (DEGs). The majority of the DEGs were identified in OPCs, the earliest cell type in the lineage, a finding resembling what we know from pyramidal neurons. The DEGs were further explored using overrepresentation analysis through WebGestalt. OPCs showed the highest number of enriched Gene Ontology (GO) terms, with two drawing particular interest, the gamma-aminobutyric acid (GABA) and the glutamate receptor (GluR) signaling pathways. The GABA signaling pathway has been associated with OPC migration, while the GluR signaling pathway is involved in the differentiation process of OPCs. SCENIC was used to analyze gene regulatory networks changes. The results indicate that EPO acts differently across the oligodendrocyte cell lineage, with enriched GO terms indicating a role not only in differentiation, but also in myelination. The results indicate that EPO acts on the oligodendrocyte lineage to support the upon EPO enhanced neuron numbers.