Aortic stiffness is associated with increased risk of cerebrovascular disease and cognitive impairments with aging, although the mechanism that contribute to this link are not well‐defined. Increases in carotid artery flow pulsatility coincide with elevated aortic stiffness and may contribute to microvascular brain damage, but studies confirming augmented flow pulsatility in the downstream microvasculature have been limited. Given the ability to assess the retinal microvasculature non‐invasively in humans and evidence that disease‐ and age‐related alterations in the retinal microvasculature mirror changes in the cerebral microvasculature, we examined the hypothesis that age‐related increases in aortic stiffness would be correlated with higher retinal arteriole flow pulsatility (systolic flow – diastolic flow / mean flow). Among 33 healthy adults (age range: 25–60 years, 47% women), aortic stiffness was assessed by carotid‐femoral pulse wave velocity (CFPWV). Retinal arteriole flow pulsatility was quantified using laser speckle flowgraphy before and during a reduction in retinal vascular resistance via 30 s of diffuse luminance flicker. Multiple regression analysis including CFPWV, mean arterial pressure (MAP), and heart rate (HR) as independent variables demonstrated that lower MAP and HR, but not higher CFPWV, were significant determinants of greater flow pulsatility at baseline (CFPWV: β=0.10, P=0.548; MAP: β= −0.40, P=0.021; HR: β= −0.50, P=0.002; model R2=0.43, P=0.001). However, after reducing retinal vascular resistance (P<0.001) via 30s of diffuse luminance flicker, multiple regression analysis demonstrated that higher CFPWV was a significant determinant of greater retinal arteriole flow pulsatility (β=0.35, P=0.026) in addition to lower MAP (β= −0.54, P=0.001) and HR (β= −0.43, P=0.005) (model R2=0.51, P<0.001). In summary, no relation was observed between higher CFPWV and retinal arteriole flow pulsatility at rest, but a positive relation was observed during arteriole vasodilation. These findings suggest that greater vascular resistance may be an important compensatory mechanism to reduce transmission of flow pulsatility.Support or Funding InformationAHA 17POST33440101, T32 HL07121 NHLBI (P01HL014388), U54 TR001356 and AHA (13DG143400012).