THE endothelial cell-specific vascular endothelial growth factor (VEGF
)(1-5) and its cellular receptors Flt-1 (refs 6,7) and Flk-1 (refs 8,9
) have been implicated in the formation of the embryonic vasculature.
This is suggested by their colocalized expression during embryogenesis
(10,11) and the impaired vessel formation in Flk-1 (ref. 12) and Flt-1
(ref. 13) deficient embryos. However, because Flt-1 also binds placen
tal growth factor(14,15), a VEGF homologue, the precise role of VEGF w
as unknown. Here we report that formation of blood vessels was abnorma
l, but not abolished, in heterozygous VEGF-deficient (VEGF(+/-)) embry
os, generated by aggregation of embryonic stem (ES) cells with tetrapl
oid embryos (T-ES)(16,17), and more impaired in homozygous VEGF-defici
ent (VEGF(-/-)) T-ES embryos, resulting in death at mid-gestation. Sim
ilar phenotypes were observed in F-1-VEGF(+/-) embryos, generated by g
ermline transmission. We believe that this heterozygous lethal phenoty
pe, which differs from the homozygous lethality in VEGF-receptor-defic
ient embryos, is unprecedented for a targeted autosomal gene inactivat
ion, and is indicative of a tight dose-dependent regulation of embryon
ic vessel development by VEGF.