Locusta-DP is a corticotropin-releasing factor (CRF)-related diuretic peptide isolated from the migratory locust Locusta migratoria. At nanomolar concentrations, synthetic Locusta-DP stimulated fluid secretion and cyclic AMP production by Malpighian tubules isolated in vitro and increased the rate of amaranth clearance in starved locusts to levels comparable with those observed during post-feeding diuresis. The peptide also caused a marked (approximately 10 %), but short-lived, reduction in the haemolymph volume of starved locusts. A polyclonal antiserum raised against Locusta-DP(29-46) was shown to block peptidergic signal transfer in vitro and in vivo. Pre-treatment of Locusta-DP (5 nmol l-1) with antiserum diluted 1:100 resulted in a rapid reduction in the free peptide concentration to less than 1 nmol l-1, the threshold for a measurable effect on cyclic AMP production by isolated tubules. In intact insects, passive immunization with Locusta-DP antiserum blocked increases in the rate of amaranth clearance in response to exogenous diuretic peptide or in response to feeding. The latter was due specifically to the binding of Locusta-DP, because when the relevant antibodies were preadsorbed with Locusta-DP(29-46), the antiserum had no effect on amaranth clearance by recently fed insects. This provides unequivocal evidence of a hormonal function for Locusta-DP in the control of primary urine production.