Previous population-based data suggest that retinal arteriolar diameter is wider in women than in men. Estrogen exposures could account for this difference. To evaluate the effects of HRT on small blood vessels, we assessed the relationship between use of hormone replacement therapy (HRT) and retinal microvascular diameter in older women in the Blue Mountains Eye Study baseline population (n = 1993, age >/= 49 years). Information on HRT use was recorded by trained interviewers. A computer-assisted program measured retinal vessel diameters from digitized photographs. Average arteriolar diameter was calculated as the central retinal arteriolar equivalent (CRAE); the lowest quintile of CRAE was considered generalized arteriolar narrowing. Of the 1897 women with complete data, 79 (4%) were premenopausal and 315 (17%), 224 (12%), and 1279 (67%) were current, past, and never HRT users, respectively. Among women aged < 65 years, multivariate-adjusted mean CRAE for the four groups was 196, 198, 201, and 200 microm (P < 0.0001), respectively. Among postmenopausal women >/= 65 years, multivariate-adjusted mean CRAE was 187, 188, 191 microm (P < 0.0001), respectively, for current, past, and never users. Current HRT users were 40% more likely than never users to have generalized arteriolar narrowing (95% confidence interval 1.0-2.1). It appears that the use of HRT in older women may not have any long-term vasodilatory effects on retinal arterioles. These data do not support our hypothesis that exogenous estrogen exposures account for observed wider retinal arteriolar diameters in women.