AbstractAllatostatins isolated from the cockroach Diploptera punctata are a family of neuropeptides that inhibit juvenile hormone synthesis in cockroaches and related insects but not in flies. In cockroaches, these widely distributed peptides have been shown to have other functions. This report provides evidence for the presence of allatostatin‐like peptides in Drosophila melanogaster by demonstration of allatostatic activity of extracts of central nervous system from larvae and adults on corpora allata of Diploptera and by immunocytochemical localization of peptides in Drosophila with monoclonal antibody against Diploptera allatostatin I. Extract of adult central nervous system showed four times more allatostatic activity than that of the larva or twice the activity per unit volume of central nervous system. This is reflected in an increase in number and arborization of immunoreactive neurons in the adult. The immunoreactive neurons in the central nervous system appear to be interneurons, with the exception of motoneurons in the last abdominal neuromere that project to muscles of the hindgut, a pair of peripheral cells in each of two thoracic segments in the larva and on nerves to wings and halteres in the adult, and endocrine cells of the midgut epithelium. Nerves to the corpus allatum were not immunoreactive. The presence of Diploptera allatostatin‐like peptides in interneurons and motoneurons, in the neurohemal networks, and in endocrine cells of the midgut and their absence in nerves to the corpus allatum in Drosophila suggests that these peptides may function as neuromodulators, myomodulators, and neurohormones and not as regulators of the corpus allatum. © 1995 Wiley‐Liss, Inc.