Besides the regulation of hematopoiesis, granulocyte-macrophage colony-stimulating factor (GM-CSF) induces the expression of a functional program in cultured endothelial cells (ECs) related to angiogenesis and to the their survival in bone marrow microenvironment. ECs express the specific GM-CSF receptor that signals through the recruitment and the activation of Janus kinase (JAK)2 (Soldi et al., Blood 89, 863-872, 1987). We now report that GM-CSF in vivo induces angiogenesis and activates JAK-2 and signal transducers and activators of transcription (STAT)-3. This cytokine has an angiogenetic activity in chick chorioallantoic membrane (CAM) without recruitment of inflammatory cells and induces vessel sprouting from chicken aorta rings. When added to CAM, subnanomolar concentrations of GM-CSF cause a rapid phosphorylation in tyrosine residues of JAK-2 persisting at least for 10 min. Furthermore, we show that signal transducers and activators of transcription (STAT)-3, but not STAT-5, also are phosphorylated for 30 min after GM-CSF stimulation. AG-490, a JAK-2 inhibitor, reduced in a dose-dependent manner the angiogenic effect of GM-CSF in CAM. These findings provide the first evidence that the JAK-2/STAT-3 pathway is activated in vivo and participates in vessel formation triggered by GM-CSF.