To understand roles of transcriptional factors and miRNAs in regulating gene expression in the epididymis from postnatal development through aging, systematic profiling of genes and miRNAs expressed in the newborn, young adult, and aged human epididymides was performed by cDNA array and miRNA array analysis, respectively. The newborn human epididymis expressed the fewest mRNAs but the largest number of miRNAs, whereas the adult and aged epididymides expressed the most mRNAs but the fewest miRNAs, a negative correlation between mRNAs and miRNA during aging. By integrative analysis, a set of miRNA targets were predicted based on the miRNA and cDNA arrays. In the newborn epididymis, 127 miRNAs were exclusively or preferentially expressed but only 3 and 2 miRNAs showed an age-enriched expression pattern in the adult and aged epididymides, respectively. This study provides a basic database as well as new insights and foundations for further studies on the complex regulation of gene expression in the epididymis.