The volume of the mushroom bodies of the brains of honey bee queens (Apis mellifera) was estimated using the method of Cavalieri. Tissue sampled was obtained from queens in five different behavioral and reproductive states: 1-day-old virgin queens, 14-day-old virgin queens, 14-day-old instrumentally inseminated queens, 9- to 13-day old naturally mated queens, and 5-month-old naturally mated queens. There were significant volume changes within the mushroom bodies during the first 2 weeks of adult life. The volume occupied by the somata of the intrinsic neuronal population (Kenyon cells) of the mushroom bodies decreased by approximately 30% and the volume of the neuropil of the mushroom bodies increased between 25 and 50%. These volume changes are strikingly similar to those previously reported to occur for worker honey bees switching from hive activities to foraging (Withers, Fahrbach, & Robinson, 1993). However, in this study they were found even in queens that had no flight experience. In addition, queens exhibiting these volume changes were found to have low blood levels of juvenile hormone, while previous studies have shown that foraging worker honey bees have high hormone levels. These results suggest that some aspect of behavioral development common to both the queen and the worker castes is fundamental to protocerebral volume changes early in adulthood in honey bees. If juvenile hormone regulates this process, results from queens suggest that it may play an organizational role.