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Objectives To investigate the roles of steroid hormones, including estrogen, progesterone, and testosterone, in the morphology and vascularization of the female bladder. Methods A total of 70 adult, female Sprague-Dawley rats were divided into 7 groups: group 1, sham; group 2, ovariectomized rats without treatment; group 3, low-dose estradiol; group 4, high-dose estradiol; group 5, progesterone; group 6, estradiol combined with progesterone; and group 7, testosterone. All were given for 4 weeks. The serum steroid hormone levels were determined by radioimmunoassay. The total weight and thickness of the bladder were measured. Morphologic changes were observed under light and electron microscopy. The expression of vascular endothelial growth factor (VEGF) in the bladder was evaluated by immmohistochemistry and Western blotting. Results The ovariectomized rats had a thinner bladder wall compared with the sham group (0.97 ± 0.11 mm vs 1.10 ± 0.10 mm, P < .05) and widened spaces between the detrusor muscle fascicles with collagen deposit. Estrogen reversed these changes, and testosterone increased the thickness of the bladder wall to 1.26 ± 0.12 mm (P < .05). VEGF staining was mainly located in the urothelium and endothelial cells, with weak staining in the smooth muscles. VEGF was almost absent in the urothelium after ovariectomy. In the estrogen- and androgen-treated groups, although the expression of VEGF was significantly greater than that in the nontreated ovariectomized group, it was still lower than normal. Conclusions Our findings suggest the importance of steroid hormones in maintaining the integrity of the bladder structure and regulating the expression of VEGF in the female urinary tract. Both estrogen and androgen can reverse the bladder muscle atrophy induced by ovariectomy. However, the decline in VEGF expression in the bladder cannot be fully recovered with either estrogen or androgen replacement.