This preliminary study used voxel- mirrored homotopic connectivity (VMHC), a novel resting-state fMRI parameter to investigate inter-hemispheric functional activity changes in restless legs syndrome (RLS). Ten RLS patients and ten age- and gender-matched healthy controls were recruited for comparison. The RLS group showed increased VMHC in the amygdala, putamen and insula, as compared to normal controls. Increased VMHC in the insular cortex and putamen might reflect their functions in perception and motor control. Increased VMHC in the amygdalae could be relevant to the disturbed sleep at night, considering its primary role in the processing of memory and emotion.