Identification of neuropeptide F in the yellow fever mosquito, Aedes aegypti
Stanek, Dawn M.
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A neuropeptide F (NPF) was isolated from an extract of adult Aedes aegypti (L.) with high performance liquid chromatography, and identified based on its immunore activity in the Drosophila NPF radioimmunoassay. Degenerate primers designed from the partial sequence of the purified peptide were used in the polymerase chain reaction to amplify products that contained the cDNA encoding Aedes aegypti NPF (AeaNPF). Similar to invertebrate NPF's and the neuropeptide Y family in vertebrates, the encoded prepropeptide was processed into an NPF of 36 residues with a carboxyl terminal amide. Immunocytochemistry and Northern analyses demonstrated NPF expression in the brain and midgut of larval and adult A. aegypti. The highest titers of NPF in the hemolymph of females were detected pre-blood meal and 24 hours post-blood meal. To determine if conservation of sequence was reflected in function, a bioassay was conducted with synthetic <i>Aea</i>NPF peptide to test its effects on trypsin levels after a blood meal. Identification and functional studies of regulatory peptides localized in the mosquito brain and midgut are crucial and may reveal opportunities to control mosquito populations and to make evolutionary comparisons to existing members of the invertebrate NPF family.