PMID:12163486

Citation	Radford, JC, Davies, SA and Dow, JA (2002) Systematic G-protein-coupled receptor analysis in Drosophila melanogaster identifies a leucokinin receptor with novel roles. J. Biol. Chem. 277:38810-7
Abstract	Leucokinins are insect neuropeptides that stimulate hindgut motility and renal fluid secretion. Drosophila has a single leucokinin gene, pp, encoding the longest known leucokinin, Drosokinin. To identify its receptor, a genome-wide scan for G-protein-coupled receptors was performed in silico and candidate receptors identified by similarity to known tachykinin receptors. The deduced peptides were expressed, with a transgene for the calcium reporter aequorin, in S2 cells and only one gene (CG10626) encoded a protein that responded to Drosokinin. The properties of the heterologously expressed receptor (action through intracellular calcium with an EC(50) of 4 x 10(-11) m and a t(1/2) <1 s) match closely those reported for the action of Drosokinin on Malpighian (renal) tubules. Antibodies raised against the receptor identified known sites of leucokinin action: stellate cells of the Malpighian tubule, two triplets of cells in the pars intercerebralis of the adult central nervous system, and additional cells in larval central nervous system. Western blots and reverse transcription-PCR confirmed these locations, but also identified expression in male and female gonads. These tissues also displayed elevated calcium in response to Drosokinin, demonstrating novel roles for leucokinin. A functional genomic approach has thus yielded the first complete characterization of a leucokinin receptor in an insect.
Links	PubMed Online version:10.1074/jbc.M203694200
Keywords	Amino Acid Sequence; Animals; Base Sequence; Calcium/metabolism; Drosophila Proteins/classification; Drosophila Proteins/genetics; Drosophila Proteins/metabolism; Drosophila melanogaster/genetics; Drosophila melanogaster/metabolism; Embryo, Nonmammalian/anatomy & histology; Embryo, Nonmammalian/physiology; Female; GTP-Binding Proteins/classification; GTP-Binding Proteins/genetics; GTP-Binding Proteins/metabolism; Genes, Insect; Genes, Reporter; Humans; Male; Malpighian Tubules/cytology; Malpighian Tubules/metabolism; Molecular Sequence Data; Nervous System/cytology; Nervous System/metabolism; Neuropeptides/metabolism; Receptors, Cell Surface/classification; Receptors, Cell Surface/genetics; Receptors, Cell Surface/metabolism; Receptors, Neuropeptide/classification; Receptors, Neuropeptide/genetics; Receptors, Neuropeptide/metabolism; Sequence Alignment; Transgenes
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