Regulatory peptides were immunolocalized in the midgut of the fruit fly Drosophila melanogaster. Endocrine cells were found to produce six different peptides: allatostatins A, B and C, neuropeptide F, diuretic hormone 31, and the tachykinins. Small neuropeptide-F (sNPF) was found in neurons in the hypocerebral ganglion innervating the anterior midgut, whereas pigment-dispersing factor was found in nerves on the most posterior part of the posterior midgut. Neuropeptide-F (NPF)-producing endocrine cells were located in the anterior and middle midgut and in the very first part of the posterior midgut. All NPF endocrine cells also produced tachykinins. Endocrine cells containing diuretic hormone 31 were found in the caudal half of the posterior midgut; these cells also produced tachykinins. Other endocrine cells produced exclusively tachykinins in the anterior and posterior extemities of the midgut. Allatostatin-immunoreactive endocrine cells were present throughout the midgut. Those in the caudal half of the posterior midgut produced allatostatins A, whereas those in the anterior, middle, and first half of the posterior midgut produced allatostatin C. In the middle of the posterior midgut, some endocrine cells produced both allatostatins A and C. Allatostatin-C-immunoreactive endocrine cells were particularly prominent in the first half of the posterior midgut. Allatostatin B/MIP-immunoreactive cells were not consistently found and, when present, were only weakly immunoreactive, forming a subgroup of the allatostatin-C-immunoreactive cells in the posterior midgut. Previous work on Drosophila and other insect species suggested that (FM)RFamide-immunoreactive endocrine cells in the insect midgut could produce NPF, sNPF, myosuppressin, and/or sulfakinins. Using a combination of specific antisera to these peptides and transgenic fly models, we showed that the endocrine cells in the adult Drosophila midgut produced exclusively NPF. Although the Drosophila insulin gene Ilp3 was abundantly expressed in the midgut, Ilp3 was not expressed in endocrine cells, but in midgut muscle.

The distribution and morphology of neurons containing allatostatin-related substances in the brain of the locust Schistocerca gregaria was investigated using an antiserum against Diploptera punctata allatostatin I (Dip-allatostatin I, APSGAQRLYGFGL-amide). In each brain hemisphere, about 550 neurons in the midbrain and 500 neurons in the optic lobe exhibit Dip-allatostatin I-like immunoreactivity, including about eight lateral neurosecretory cells with processes to the retrocerebral complex. All major brain areas except the antennal lobe, the mushroom body, and large parts of the lamina, are innervated by Dip-allatostatin I-immunoreactive processes. Immunostaining in the central complex was studied in detail. The central complex is innervated by more than 260 Dip-allatostatin I-immunoreactive neurons belonging to six different cell types, four sets of tangential neurons and two sets of columnar neurons. These neurons give rise to intense immunostaining in the protocerebral bridge, in several layers of the upper division of the central body, and in the dorsalmost layer of the lower division of the central body. Double-label experiments show colocalization of Dip-allatostatin I- and serotonin-like immunoreactivities in one type of columnar and one type of tangential neurons of the central complex. The similar patterns of Dip-allatostatin I- and galanin message-associated peptide-like immunoreactivities result from cross-reactivity of the anti-galanin message-associated peptide antiserum with Dip-allatostatin I. The results provide further insight into the anatomical and neurochemical organization of the locust central complex and suggest a prominent neuroactive role for Dip-allatostatin I-related peptides in this brain area.

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Adipokinetic hormone (AKH) peptides in insects serve the endocrine control of energy supply. They also produce, however, neuronal, vegetative, and motor effects, suggesting that AKHs orchestrate adaptive behavior by multiple actions. We have cloned, for Periplaneta americana, the AKH receptor to determine its localization and, based on current measurements in neurons and heterologous expression systems, the mechanisms of AKH actions. Apart from fat body, various neurons express the AKH receptor, among them abdominal dorsal unpaired median (DUM) neurons, which release the biogenic amine octopamine. They are part of the arousal system and are involved in the control of circulation and respiration. Both the two Periplaneta AKHs activate the Gs pathway, and AKH I also potently activates Gq. AKH I and--with much less efficacy--AKH II accelerate spiking of DUM neurons through an increase of the pacemaking Ca2+ current. Because the AKHs are released from the corpora cardiaca into the hemolymph, they must penetrate the blood-brain barrier for acting on neurons. That this happens was shown electrophysiologically by applying AKH I to an intact ganglion. Systemically injected AKH I stimulates locomotion potently in striking contrast to AKH II. This behavioral difference can be traced back conclusively to the different effectiveness of the AKHs on the level of G proteins. Our findings also show that AKHs act through the same basic mechanisms on neuronal and nonneuronal cells, and they support an integration of metabolic and neuronal effects in homoeostatic mechanisms.