Urban songbirds of several species more vigorously defend their territories in response to conspecific song playback than do their rural counterparts, but the hormonal basis of this behavioral difference is unclear. It is well established in vertebrates that both testosterone and corticosterone affect the intensity of territoriality. Previous studies have found no evidence that initial (i.e., immediately following territorial challenge, but prior to restraint) plasma testosterone accounts for the elevated territorial aggression of urban birds. Determining if testosterone still contributes to urban-rural differences in territoriality requires also assessing males' abilities to transiently increase plasma testosterone (in response to an injection of gonadotropin-releasing hormone). We tested whether these hormones are correlated with the territorial response to conspecific song playback in urban and rural male Song Sparrows (Melospiza melodia) in Montgomery County, Virginia. We found that the elevated territorial aggression of urban sparrows was not related to variation in either initial plasma testosterone or the ability to transiently increase testosterone. In contrast, despite no overall habitat difference in initial corticosterone, levels of this hormone were positively correlated with territoriality in urban and rural sparrows. Furthermore, for a given level of corticosterone, urban sparrows were more territorially aggressive. Our findings suggest that initial corticosterone may either play a role in the regulation of persistent differences in territorial behavior between free-ranging urban and rural male Song Sparrows or be affected by the intensity of behavioral response to territorial challenge.