The midgut of insects has recently been shown to contain numerous endocrine-like cells and the midgut is now considered one of the largest endocrine organs in the insect. Using immunohistochemistry, radioimmunoassay, and muscle bioassay techniques, the midgut of the adult locust, Locusta migratoria, has been investigated for the distribution and possible function of FMRFamide-related peptides contained within these endocrine-like cells and innervation. Endocrine-like cells containing RFamide-like immunoreactivity were observed to be unequally distributed throughout the midgut. RFamide-like immunoreactivity was also seen in the ingluvial ganglion and in the nerves projecting posteriorly to the midgut. These axonal tracts resulted in extensive arborizations over the posterior midgut which were RFamide-like immunoreactive. Radioimmunoassay indicated larger amounts of FMRFamide equivalents in female locust midgut as compared to males with an unequal distribution of FMRFamide-like immunoreactivity in the gastric caeca and in the anterior and posterior parts of the midgut. Circular muscle contraction of the midgut was monitored using a ring-type preparation. Structure/activity studies have shown that the only FMRFamide-related peptides tested that alter circular muscle contraction of the midgut are those that belong to the subfamily referred to as myosuppressins. SchistoFLRFamide, leucomyosuppressin, and ManducaFLRFamide were each capable of lowering basal tonus and inhibiting spontaneous and proctolin-induced contractions of midgut muscle. Further structure/activity studies indicated that HVFLRFamide is the minimum sequence required to achieve inhibition comparable to the parent compound. This work suggests that a possible function for the FMRFamide-related peptides contained within the endocrine cells and innervation of the midgut of the locust may be in modulating midgut contraction and thereby playing a role in digestion.