Insulin-like peptide DILP6 is a component of the insulin/insulin-like growth factor signalling pathway of Drosophila. Juvenile hormone (JH) and dopamine (DA) are involved in the stress response and in the control of reproduction. In this study, we investigate whether DILP6 regulates the JH and DA levels by studying the effect of a strong hypomorphic mutation dilp6(41) on JH and DA metabolism in D. melanogaster females. We show that DILP6 regulates JH and DA metabolism: the mutation dilp641 results in a reduction in JH-hydrolysing activity and an increase in the activities of DA synthesis enzymes (alkaline phosphatase (ALP) and tyrosine hydroxylase (TH)). In the mutant females, we also found increased fecundity in addition to the intensity of the response (stress reactivity) of ALP and TH to heat stress. As we showed previously, this suggests an increased level of JH synthesis. We confirm this suggestion by treating the mutant females with the JH inhibitor, precocene, which restors the activity and stress reactivity of ALP and TH as well as fecundity to levels similar to those in the control flies. The data suggest a feedback system in the interaction between JH and DILP6 in which DILP6 negatively regulates the JH titre via an increase in the hormone degradation and a decrease in its synthesis. (C) 2016 Elsevier Inc. All rights reserved.