Allatostatins are neuropeptides that inhibit the production, by the corpora allata, of a major insect hormone, juvenile hormone. These peptides are produced by cells of the brain and ganglia as well as by midgut endocrine cells. Transport from these sites may contribute to the allatostatin content in the hemolymph (insect blood). Using a monoclonal antibody against Diploptera punctata allatostatin I (A-P-S-G-A-Q-R-L-Y-G-F-G-L-NH2) and in situ hybridization with a digoxigenin-labeled cRNA probe generated from a portion of the allatostatin gene, it is demonstrated that allatostatin is present in and synthesized by granular hemocytes of D. punctata. About 5% of the hemocytes react with anti-allatostatin antibody and a similar number hybridize with a cRNA probe that detects allatostatin-specific mRNA. Electron micrographs showed that allatostatin-immunoreactive material occurs in membrane-bound, uniformly dense granules that frequently fill fusiform-shaped cells. Allatostatin in cell and plasma fractions of hemolymph quantified by enzyme-linked immunosorbent assay and by bioassay for inhibition of juvenile hormone synthesis in vitro indicated that about equal quantities (0.1-0.2 fmol/microl) are present in cell and plasma fractions. The production of allatostatin by hemocytes suggests that allatostatins may function as regulatory peptides in hemolymph activities in addition to their other known functions.
The identification of neuropeptides that inhibit juvenile hormone (JH) synthesis by the corpora allata (CA) has verified the existence of these allatostatins, which, from much experimental evidence, have long been postulated to occur. It also makes possible new approaches for studying the role of allatostatins in the regulation of JH synthesis. Allatostatins, localized immunocytochemically, occur in lateral neurosecretory cells of the brain that innervate the CA. Presumably their effect on the CA results from the release of allatostatins at these nerve endings. Allatostatins also occur in the hemolymph in cockroaches and have been shown to act on the CA through this pathway. The ability of allatostatins to inhibit CA depends not only on the concentration of the peptides but also on the sensitivity of the CA to them. Male Diploptera punctata were treated with JH analog following denervation of CA and implanted with a previtellogenic or vitellogenic ovary or injected with saline. Animals implanted with a vitellogenic ovary, compared to the previtellogenic ovary or saline, showed significantly increased JH synthesis by their CA and a reduced amount of allatostatin in the hemolymph. The denervated CA from these JH analog treated animals, following implantation with a previtellogenic and vitellogenic ovary, showed a tendency toward increased and decreased sensitivity, respectively, to a given dose of allatostatin in vitro compared to those from saline injected controls. Experiments such as these suggest that changes in release of allatostatins and in sensitivity of CA to them could be postulated to be major factors regulating JH synthesis in the cockroach.
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