The angiogenic effects of 17β-oestradiol (E₂) in the mouse endometrium are mediated by vascular endothelial growth factor-A (VEGFA). We analysed the temporal and spatial changes in VEGFA isoform and (co)receptor expression in ovariectomised mouse uteri following E₂treatment. VEGFA isoform and receptor mRNA were quantified in whole uterine tissue collected 2, 6, 12 and 24 h after E₂or vehicle treatment. Laser capture microdissection was used to investigate mRNA expression in epithelial, stromal and myometrial tissues separately. Endothelial cell proliferation, VEGFA and VEGF receptor-2 (VEGFR2) protein were visualised using immunohistochemistry. Endometrial endothelial cell proliferation was only observed 24 h after E₂treatment. In whole uterine tissue, totalVegfa,Vegfa₁₆₄andVegfa₁₂₀mRNA expression increased 2 h post E₂treatment, and then decreased by 24 h.Vegfa₁₈₈expression was lower in E₂-treated animals at all time points relative to control animals.Vegfr2and neuropilin-1 (Nrp1) mRNA expression did not change following E₂treatment;Nrp2expression decreased by 24 h. When uterine compartments were considered separately at 24 h post E₂or vehicle, stromalVegfa₁₂₀,Vegfa₁₈₈andVegfr2mRNA expression and myometrialVegfa₁₂₀andVegfa₁₈₈mRNA expression were reduced in E₂-treated mice relative to controls, whereas epithelialVegfa₁₈₈mRNA expression increased. The highest VEGFA immunoexpression was observed in luminal epithelium; expression increased at 24 h relative to other time points. No changes were noted in VEGFR2 immunoexpression among treatment groups. We have provided the first evidence that VEGFA isoform and receptor mRNA expression are differentially regulated by E₂in different uterine cell compartments.