The sea lamprey (Petromyzon marinus L.) represents one of the two most ancient classes of vertebrates and possesses a functional hypothalamus-pituitary-gonadal axis. However, the presence and functionality of androgens in the sea lamprey remain elusive. Recently, 15alpha-hydroxytestosterone (15alpha-T) has been found in sea lamprey gonads and blood plasma. In this study we examined changes of circulatory concentrations of 15alpha-T in response to gonadotropin releasing hormone (GnRH) treatments. Plasma concentrations of 15alpha-T in sea lamprey increased 2-5 times for all GnRH-injected sea lamprey compared to controls (P < 0.001). However, there were no differences among responses: (1) to the two forms of GnRH (lamprey GnRH I or lamprey GnRH III), (2) to the doses delivered (50, 100, or 200 microg/kg), or (3) between post-injection sample intervals (8 or 24 h). Between lampreys from the Atlantic Ocean and Great Lakes sites, two of seven GnRH form and dosage comparisons showed between-site differences, but were not believed to represent an overall between-site difference. These are the first data to show a response of a C19 steroid to GnRH stimulation in sea lamprey.