Accumulation of neuropeptides in the cerebral neurosecretory system of Manduca sexta Larvae Parasitized by the Braconid Wasp Cotesia congregata

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Abstract
Fifth in star larvae of Manduca sexta that were parasitized by the braconid wasp Cotesia congregata failed to develop after the parasitoid larvae emerged, and these host larvae lingered for 2–3 weeks in a quiescent, nonfeeding state without initiating a larval molt or metamorphosis. This study was focused on the neuroendocrine changes associated with the host's developmental arrest. Immunohistochemical studies suggested that the host brain neurosecretory cells as well as their axon terminals in the corpora cardiaca-corpora allata complex accumulated multiple neuropeptides. The extent of accumulation in cells and axons increased with time, so that hosts examined 7–14 days after the wasps emerged showed the most intense staining with antibodies against prothoracicotropic hormone, bombyxin, allatotropin, allatostatin, diuretic hormone, eclosion hormone, proctolin, and FMRFamide. Increased levels of prothoracicotropic hormone and FMRFamide-like peptides in the brains of parasitized larvae were confirmed using Western blots and enzyme-linked immunosorbent assay (ELISA), respectively. Starvation of the unparasitized larvae induced some accumulation of the neuropeptides; however, the intensity of staining and number of immunopositive cells and axons were in most cases clearly higher in the parasitized larvae. Our results suggest that accumulation of the neuropeptides is associated with developmental arrest of parasitized larvae. Because a similar developmental arrest occurs in a wide range of parasitized insects, our findings may have relevance for many other species. Moreover, these data illustrate the potential value of using parasitized M. sexta larvae as a model for studying the mechanisms governing the rates of neuropeptide expression, processing, packaging, and release, as well as providing a rich source of neuropeptides, thus facilitating their isolation and characterization.

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