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Although Neuropeptide F (NPF) has been identified in different insect species, its function has mainly been studied in the fruit fly, Drosophila melanogaster, where it regulates diverse physiological processes, such as learning, stress responses and male courtship behavior. In locusts, only a truncated form of the "full-length" NPF (the biologically active "trNPF") has been isolated. This 9 AA peptide stimulates oocyte maturation, food intake and weight increase in adult desert locusts (Schistocerca gregaria [Forskal]). In this study, we investigated whether this peptide is also involved in the regulation of male reproductive physiology in this orthopteran species. Daily injections of trNPF in adult males resulted in proportionally heavier testes and seminal vesicles, while RNAi-mediated knockdown of the Schgr-NPF precursor transcript gave rise to proportionally lighter testes and seminal vesicles. Furthermore, adult males precociously displayed courtship behavior when injected daily with trNPF, while this behavior was inhibited or delayed by RNAi knockdown of the Schgr-NPF precursor transcript. In order to further analyze these effects of trNPF on male reproductive physiology, fertility of males was tested by analyzing progeny numbers following copulation with untreated females. In this way, we showed that daily trNPF injection in adult males resulted in a larger egg pod size and a higher percentage of hatched eggs per egg pod after copulation, while RNAi knockdown caused the opposite effects. Taken together, we provide clear evidence for a role of trNPF in the regulation of reproductive physiology in adult males of the desert locust, S. gregaria. Possible modes of action of trNPF in influencing these reproductive processes in male locusts are discussed. (C) 2013 Elsevier Ltd. All rights reserved.