Primary monosymptomatic nocturnal enuresis (PMNE) is a common disorder in school-aged children. However, little is known about resting-state neural function in individuals with PMNE. In this work, resting-state functional magnetic resonance imaging (fMRI) was used to investigate changes in spontaneous brain activity in children with PMNE. We analyzed resting-state fMRI data using statistical parametric mapping (SPM) and a resting-state fMRI data analysis toolkit (REST). Regional homogeneity (ReHo) and amplitude of low-frequency fluctuation (ALFF) values were calculated to analyze spontaneous brain activity in 16 children with PMNE and 16 healthy controls. Children with PMNE exhibited significant differences in ALFF or ReHo in the left inferior frontal gyrus, medial frontal gyrus (Brodmann area, BA 10), and left midbrain. Abnormalities in BA 10 and the inferior frontal gyrus may affect children's decision-making with regard to voiding; abnormalities in the midbrain in PMNE children may influence the internal signal transmission in their bladder control network. Our data indicate that, in children with PMNE, several brain areas related to the micturate control network undergo developmental delay. Neurourol. Urodynam. 31:99–104, 2012. © 2011 Wiley Periodicals, Inc.