Diploptera punctata allatostatins are brain neuropeptides that inhibit juvenile hormone synthesis by corpora allata. They also occur in nerves of many organs other than corpora allata. The distribution of allatostatins in, and the effect of allatostatins on two other organs, antennal pulsatile organ and hindgut, are demonstrated here. Allatostatin I-like immunoreactive material is present in cells of subesophageal and terminal abdominal ganglia; these ganglia are known to contain the cells that project to antennal heart nerve and proctodeal nerve, respectively. Electron micrographs of both organs show nerve terminals with allatostatic immunoreactive granules along with terminals containing nonimmunoreactive granules. Immunoreactive neurosecretory granules are about 200 nm in largest dimension. In the antennal pulsatile organ, profiles of the nerve terminals are larger in the ampullar wall than in the muscle; in hindgut the terminals with immunoreactive granules are associated with the muscle net below the circular muscle. Hindgut responded to allatostatins I and IV with a dose-dependent decrease in amplitude and frequency of contraction that was reversible, with the threshold concentration for response between 10(-8) and 10(-7) M. In contrast, pulsatile organ muscle showed no such change with either allatostatin at 10(-7)-10(-4) M. However, both organs responded to proctolin with increased amplitude and frequency of contractions. Allatostatins I and IV inhibited the proctolin-induced increase of hindgut contraction, whereas no such effect was seen in antennal pulsatile organ muscle. Extract of antennal pulsatile organ muscle showed proctolin-like bioactivity that comigrated with authentic proctolin on three sequential HPLC systems.