Based on previous evidence suggesting abnormalities in the brain microvasculature, we examined basement membrane collagen in isolated cerebral microvessels (CMV) from subjects with Alzheimer's disease (AD) and age-matched controls. Concentrations of hydroxyproline, the principal constituent of collagen IV, were significantly increased by 55% in CMV from AD subjects compared to controls. This result was corroborated by the finding of 60% increased total collagen content in CMV as evident by the selective binding of Sirius red dye. Hydroxyproline and collagen concentrations in samples of cerebral cortex assayed in parallel were 6-20 times smaller than in CMV and were not changed between controls and AD subjects. To further differentiate AD and control samples, fractions of CMV were solubilized and the pepsin digested collagen proteins resolved by SDS-PAGE. Upon immunoblotting, AD samples with increased collagen revealed proportionally greater specific immunoreactivities detected by antibodies to collagen IV. Our observations suggest altered collagen IV content of cerebral vessels in subjects with AD that may affect brain microvascular functions.