The basis for the steroidogenic differences between the human fetal adrenal (HFA) and adult adrenal is not well defined. However, gene expression clearly plays a critical role in defining their distinct steroidogenic and structural phenotypes. We used DNA microarrays to compare expression levels of several thousand transcripts between the HFA and adult adrenal gland. Gene profiling was done using seven independent microarrays that contained between 7075 and 9182 cDNA elements. Twenty-five transcripts were found to have a greater than 5-fold difference in expression between HFA and adult adrenals. The largest differences were observed for transcripts that encode insulin-like growth factor-II (IGF-II) (25-fold higher in HFA) and 3beta hydroxysteroid dehydrogenase (3betaHSD) (21-fold higher in adult). The vast majority of the 25 transcripts have not been studied with regard to adrenal function. We also determined the transcripts that had the highest signal intensities, which is an approximate measure of expression level, for both the fetal and adult adrenal RNA samples. The enzyme 24-dehydrocholesterol reductase, which is involved in cholesterol biosynthesis, exhibited the highest signal intensity for fetal adrenal RNA. For adult adrenal mRNA, the expression of 11beta-hydroxylase transcripts was found to have the highest signal intensity ranking. Overall, 10 of the top 20 highest signal intensities were similar for adult and fetal adrenal transcripts. The gene profile data for fetal vs. adult adrenal glands should provide valuable information that could help define mechanisms involved in adrenal growth and development.