Previous studies have yielded conflicting results, whether extracellular adenosine generation and signaling contributes to hypoxia-induced increases in renal erythropoietin (EPO) secretion. Here, we combined pharmacological and genetic approaches to elucidate a potential contribution of extracellular adenosine to renal EPO release in mice. To stimulate EPO secretion we used murine carbon monoxide exposure (400 and 750 ppm CO, 4h), ambient hypoxia (8% oxygen, 4h), or arterial hemodilution. As the ecto-5-nucleotidase (CD73, conversion of AMP to adenosine) is considered the pace-maker of extracellular adenosine generation, we first tested the effect of blocking extracellular adenosine generation with the specific CD73-inhibitor APCP or by gene-targeted deletion of cd73. These studies showed that neither APCP-treatment nor targeted deletion of cd73 resulted in changes of stimulated EPO mRNA or serum levels although the increases of adenosine levels in the kidney following CO exposure are attenuated in mice with APCP treatment or in cd73^-/- mice. Moreover, pharmacological studies using specific inhibitors of individual adenosine receptors (A₁AR: DPCPX; A_2AAR: DMPX; A_2BAR: PSB115; A₃AR: MRS1119) showed no effect on stimulated increases of EPO mRNA or serum levels. Finally, stimulated EPO secretion was not attenuated in gene-targeted mice lacking A₁AR^-/-, A_2AAR^-/-, A_2BAR^-/-or A₃AR^-/-. Taken together, these studies combine genetic and pharmacological in vivo evidence that increases of EPO secretion during limited oxygen availability are not affected by extracellular adenosine generation or signaling.