INTRODUCTION: Iron deficiency is a common nutritional disorder. The effect of iron deficiency on myelination of specific brain regions and their relevant behavior has not been well documented. OBJECTIVE: The objective of this study was to determine the consequences of perinatal iron deficiency on behavioral and myelination of specific brain regions in rats. METHODS: Sixteen dams were randomly assigned to iron-sufficient or low-iron diets during gestation and lactation. Thereafter, all offspring were fed the iron-sufficient diet and were assessed for behavioral and neurologic developments. Behavioral assessments included sensorimotor function tests, a recognition memory task, and a spatial learning task. The density of myelination around the hippocampus was measured by 2`,3`-cyclic nucleotide 3`-phosphohydrolase (marker of oligodendrocyte) by means of immunohistochemistry and quantified by analysis of integrated optical density. The regions of interest included the corpus callosum and the fimbria of the hippocampus. RESULTS: Iron-deficient rats showed behavioral impairments in sensorimotor functions and recognition memory task but no significant differences were found in the spatial learning task. Iron-deficient rats had significantly reduced density of myelination than control rats in the corpus callosum but had no difference in the fimbria of the hippocampus. CONCLUSIONS: This study shows that perinatal iron deficiency can significantly alter the behavioral outcomes in rat pups and can profoundly influence the development of myelination in specific brain regions.