Epidemiological evidence demonstrates an association between smallness at birth and an increased risk of cardiovascular disease in later life. In the rat, protein restriction during pregnancy results in hypertension and insulin resistance in the offspring. The aim of this study was to assess the effect of protein restriction during pregnancy on maternal vascular physiology and that of both the F₁ and F₂ offspring. Female Wistar rats were mated and fed either a control (C, 18% casein) or protein restricted (PR, 9% casein) diet from conception until term (21.5 days). The vascular reactivity of mesenteric arteries was assessed using the wire myograph in a subgroup of near-term pregnant dams, the male F₁ offspring, pregnant female F₁ offspring and male and female F₂ offspring. Systolic blood pressure was assessed using tail cuff plethysmography and eNOS mRNA levels assessed by real time PCR (TaqMan). In protein restricted pregnant dams, maternal vascular function was impaired compared to controls, and was associated with an impaired production of NO to ACh stimulation (p<0.05). Systolic blood pressure was elevated and endothelium-dependent and -independent vasodilatation was impaired in male offspring of protein restricted dams demonstrated compared to the controls (p<0.05). Pregnant female offspring from protein restricted dams displayed impaired endothelium-dependent vasodilatation near term. Male and female F₂ offspring, from both male and female parents which were protein restricted in utero, displayed elevated systolic blood pressure, which was associated with altered vascular reactivity (p<0.05). This study demonstrates that protein restriction during pregnancy alters maternal cardiovascular function and vascular function in the offspring, which persists into pregnancy and can be passed to the second generation offspring in the absence of additional dietary challenges.