Erythropoietin promotes the differentiation of fetal neural stem cells into glial cells via the erythropoietin receptor‐β common receptor/Syne‐1/H3K9me3 pathway

• Published in: CNS neuroscience & therapeutics Vol. 28; no. 9; pp. 1351 - 1364
• Main Authors: Yang, Zhen‐Hong, Zhang, Si‐Jia, Zhao, Hai‐Ping, Li, Fang‐Fang, Tao, Zhen, Luo, Yu‐Min, Wang, Rong‐Liang
• Format: Journal Article
• Language: English
• Published: Oxford John Wiley & Sons, Inc 01.09.2022; John Wiley and Sons Inc
• Subjects: Antibodies; Cell culture; Cell differentiation; Cell surface; Cell viability; Epidermal growth factor; Erythropoietin; Fetuses; Flow cytometry; Gene expression; Glial cells; Glucose; Hypoxia; Immunofluorescence; Immunoprecipitation; Ischemia; Laboratory animals; Mass spectrometry; Mass spectroscopy; neural stem and progenitor cells; Neural stem cells; Neurogenesis; Neuronal-glial interactions; Original; Performance evaluation; Progenitor cells; Proteins; Scientific imaging; Statistical analysis; Stem cells; Western blotting; β common receptor
• ISSN: 1755-5930; 1755-5949
• DOI: 10.1111/cns.13876

Aims To investigate the effect of erythropoietin (EPO) on the differentiation of neural stem cells (NSCs)/neural progenitors (NPs) in the treatment of hypoxic–ischemic injury and its potential mechanisms. Methods Fetal NSCs/NPs were treated with EPO after oxygen and glucose deprivation/reoxygenation (OGD/R). Cell viability, proliferation, and differentiation of NSCs/NPs were detected by CellTiter‐Glo, Edu assay, flow cytometry, and quantitative real‐time PCR (qPCR). Immunofluorescence staining, co‐immunoprecipitation (Co‐IP), and western blotting were used to test the existence of EPO receptor/β common receptor (EPOR/βCR) heterodimer on NSCs/NPs and the possible pathway. Results EPO treatment at different time points increased cell viability without affecting proliferation. EPO treatment immediately after OGD/R promoted oligodendrocyte and astrocyte differentiation, while decreasing neuronal differentiation of NSCs/NPs. EPOR/βCR heterodimer existed on the cell surface of the fetal cortical NSCs/NPs, EPO treatment significantly increased the mRNA expression of βCR and elevated the correlation between EPOR and βCR levels. In addition, mass spectrometry analysis identified Syne‐1 as a downstream signaling molecule of the EPOR/βCR heterodimer. Immunofluorescence staining and western blotting indicated that the βCR/Syne‐1/H3K9me3 pathway was possibly involved in the differentiation of fetal neural stem cells into the glial cell effect of EPO. Conclusion EPO treatment immediately after OGD/R could not facilitate fetal NSCs/NPs neurogenesis but promoted the formation of the EPOR/βCR heterodimer on fetal NSCs/NPs, which mediates its function in glial differentiation. EPO activates the EPOR‐βCR/Syne‐1/H3K9me3 signaling pathway and controls the cell fate switch of NSCs/NPs.