Role of H2O2 and heme-containing O2 sensors in hypoxic regulation of tyrosine hydroxylase gene expression

In the current study, we investigated links between O-2-regulated H2O2 formation and the hypoxic induction of mRNA for tyrosine hydroxylase (TK), the rate-limiting enzyme in catecholamine synthesis, in O-2-sensitive PC-12 cells. During exposure of PC-12 cells to 5% O-2, H2O2 concentration decreased by 40% as measured with 2',7'-dichlorofluorescein (DCF). Treatment with H2O2 reduced TH mRNA during normoxia and prevented the induction of TH mRNA during hypoxia. Treatment with catalase or N-(2-mercaptopropionyl)-glycine, a reducing antioxidant agent that decreases H2O2 concentration, also induced TH mRNA. Deferoxamine (DF), an iron chelator, failed to affect H2O2 formation but induced TH mRNA in normoxia and hypoxia. CoCl2 led to a decrease in H2O2 at 20 h of treatment but induced TH mRNA during normoxia and hypoxia before it affected H2O2. In conclusion, TH gene expression correlates inversely with H2O2 formation. DF and Co2+ Seem to affect TH gene expression in the mechanism downstream from the H2O2 formation rather than by interfering with the H2O2-generating activity of the O-2 sensor.