Abstract--Deficiencies in maternal endothelial NO synthase (eNOS) have been associated with pregnancy complications, intrauterine growth retardation, and altered vascular function in offspring. In the present study, we investigated the influence of the maternal eNOS genotype on offspring’s blood pressure, heart rate, and locomotor activity. The effect of maternal eNOS genotype was made by comparing telemetered blood pressure and activity between 2 groups of 13- to 16-week-old male heterozygous eNOS knockout mice, 1 produced by a cross of eNOS knockout (eNOS^-/-) mothers and wild-type (eNOS^+/+) fathers (eNOS^+/-MAT offspring, N=11), the other by a cross of eNOS^+/+ mothers and eNOS^-/- fathers (eNOS^+/-PAT offspring, N=10). Data were also collected for homozygous eNOS^-/- and eNOS^+/+ mice (N=15 each). Heterozygous eNOS knockout mice exhibited blood pressures that were intermediate to the eNOS^+/+ and eNOS^-/- groups. Relative to eNOS^+/-PAT mice, eNOS^+/-MAT mice exhibited significant increases in nocturnal diastolic arterial pressure and diurnal variations (dark-light difference) in systolic, mean, and diastolic arterial pressure. In addition, indices of spontaneous nocturnal locomotor activity, including both the proportion of time spent active and the intensity of activity when active, were also significantly increased. Heart rate did not differ between the groups. Our results suggest that the maternal eNOS genotype influences both blood pressure and behavior of offspring, possibly as a consequence of developmental programming associated with intrauterine growth retardation.