The effect of five daily intraperitoneal (i.p.) injections of NT69L on in vitro dopamine release, uptake, and [(3)H]NT binding in rat striatal tissue was investigated. NT69L perfusion increased K(+)-evoked and electrically evoked [(3)H]DA release. NT receptor-1 antagonist SR48692 inhibited the stimulatory effect of NT69L on K+-evoked [(3)H]DA release, but not on electrical depolarization. Pretreatment with NT69L, in vivo, daily for 5 days, did not cause significant change in K(+) evoked [(3)H]DA release, but reduced electrically evoked [(3)H]DA release induced by NT69L perfusion. Repeated perfusion with NT69L in vitro caused marked reduction on K(+)-evoked [(3)H]DA release and no change in electrically evoked [(3)H]DA release. [(3)H]NT binding was not significantly changed by one injection but was decreased after five injections of NT69L. Desensitization to the effects of NT69L in vitro was different depending upon whether tissue was preexposed to the compound in vivo or in vitro. These results provide further proof for the involvement of different NT receptor subtypes in mediating the effect of NT69L on dopamine release evoked by K(+) or electrical depolarization.