Erythropoietin (EPO) is established as a major regulator of erythropoiesis. However, we and others have shown that neurons express erythropoietin receptor (EPO-R), that astrocytes produce EPO and that EPO may act as a neurotrophic factor in the CNS. We also found that EPO production is activated by insulin and insulin-like growth factors (IGFs) in astrocytes in a dose-dependent manner and that IGF-I was the most potent activator. The concentrations required for half-maximal activation were 3 nM IGF-I, 10 nM IGF-II and 100 nM insulin. The oxygen concentration regulates EPO production; hypoxia stimulates EPO production in astrocytes. The stimulatory effect of IGFs and insulin on EPO production in astrocytes was not affected by the oxygen concentration of astrocyte culture. Insulin and IGFs did not increase the total protein synthesis of astrocytes but increased EPO mRNA levels, indicating that EPO production is stimulated at the mRNA level. It appeared that the growth factor-induced accumulation of EPO mRNA in astrocytes was caused by activation of the tyrosine kinase-signal transduction pathway, because tyrosine phosphorylation of receptors for IGF-I and insulin was activated when astrocytes were stimulated by these growth factors.