Cloning, Characterization, and Expression of a G-Protein-Coupled Receptor from<i>Lymnaea stagnalis</i>and Identification of a Leucokinin-Like Peptide, PSFHSWSamide, as Its Endogenous Ligand

Cox, Kingsley J. A.; Tensen, Cornelis P.; Schors, Roel C. van der; Li, Ka Wan; Heerikhuizen, Harm van; Vreugdenhil, Erno; Geraerts, W.P.M.; Burke, Julian F.

doi:10.1523/jneurosci.17-04-01197.1997

Cited by 73 publications

(71 citation statements)

References 41 publications

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“…The protein sequences of the Lymnaea stagnalis L. (Cox et al, 1997) and Boophilus microplus Canestrini (Holmes et al, 2003) leucokinin-like receptors were also used to confirm the sequence match. The BLOSUM62 matrix (default settings) was used for all BLAST analysis as above.…”

Section: Methodsmentioning

confidence: 99%

“…The protein sequences of the known leucokinin-like receptors, the Drosophila LKR (CG10626; Radford et al, 2002), the lymnokinin receptor (GenBank accession AAD11810; Cox et al, 1997), the B. microplus receptor (AAF72891; Holmes et al, 2003) and the putative A. stephensi receptor (Fig.·3) were aligned using the CLUSTAL X program (Thompson et al, 1994). The sequence alignment was annotated using BioEdit (Hall, 1999) (Fig.·4).…”

Section: Alignment and Comparison Of The Known Leucokinin Receptorsmentioning

confidence: 99%

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Functional characterisation of theAnophelesleucokinins and their cognate G-protein coupled receptor

Radford

Terhzaz

Cabrero

et al. 2004

Journal of Experimental Biology

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Drosophila leucokinin was also found to activate the Anopheles receptor with a similar EC 50 value to Anopheles leucokinin I. However, when the Anopheles peptides were applied to the Drosophila receptor, only Anopheles leucokinin I and II elicited a rise in [Ca 2+ ] i . This suggests that the Anopheles receptor has a broader specificity for leucokinin ligands than the Drosophila receptor.Antisera raised against the Anopheles receptor identified a doublet of approx. 65 and 72·kDa on western blots, consistent with the presence of four N-glycosylation sites within the receptor sequence, and the known glycosylation of the receptor in Drosophila. In Anopheles tubules, as in Drosophila, the receptor was localised to the stellate cells.Thus we provide the first identification of Anopheles mosquito leucokinins (Anopheles leucokinins) and a cognate leucokinin receptor, characterise their interaction and show that Dipteran leucokinin signalling is closely conserved between Drosophila and Anopheles.

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Section: Methodsmentioning

confidence: 99%

Section: Alignment and Comparison Of The Known Leucokinin Receptorsmentioning

confidence: 99%

Functional characterisation of theAnophelesleucokinins and their cognate G-protein coupled receptor

Radford

Terhzaz

Cabrero

et al. 2004

Journal of Experimental Biology

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“…They were first isolated from the cockroach Leucophaea maderae as a family of small peptides that stimulated hindgut motility (2)(3)(4)(5)(6). They are now known to be present in many other insect species as well as parasitic nematodes, crustaceans, and molluscs (7)(8)(9)(10)(11)(12)(13)(14)(15) (Table I). The leucokinins vary from 6 to 15 amino acids in length and are characterized by a C-terminal pentapeptide motif (FXXWG-amide) essential for biological activity.…”

mentioning

confidence: 99%

Systematic G-protein-coupled Receptor Analysis inDrosophila melanogaster Identifies a Leucokinin Receptor with Novel Roles

Radford

Davies

Dow

2002

Journal of Biological Chemistry

172

195

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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 ؋ 10 ؊11 M and a t1 ⁄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.

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“…Indeed, although the first leucokinins were characterized in an insect (Holman et al, 1984), the first gene for a leucokinin receptor, and its cognate peptide, were identified in the pond snail Lymnaea stagnalis in a single, thoroughly impressive paper (Cox et al, 1997). Similarly, leucokinin signalling is known to occur in the Acari (mites and ticks) (Holmes et al, 2000).…”

Section: Rnai -Does It Promote 'Target' Species To 'Model' Status?mentioning

confidence: 99%

Integrative physiology, functional genomics and the phenotype gap: a guide for comparative physiologists

Dow

2007

Journal of Experimental Biology

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SUMMARY Classical, curiosity-led comparative physiology finds itself at a crossroads. Major funding for classical physiology is becoming harder to find,as grant agencies focus on more molecular approaches or on science with more immediate strategic value to their respective countries. In turn, this shift in funding places Zoology and Animal Science departments under enormous stress: student numbers are buoyant, but how can research funding be maintained at high levels? Our research group has argued for the redefinition of integrative physiology as the investigation of gene function in an organotypic context in the intact animal. Implicit in this definition is the use of transgenics and reverse genetics to manipulate gene function in a cell-specific manner; this in turn implies the use of a genetically tractable `model organism'. The significance of this definition is that it aligns integrative physiology with functional genomics. Again, functional genomics draws heavily on reverse genetics to elucidate the function of novel genes. The phenotype gap (the mismatch between what a genetic model organism's genome encodes and the reasons that it has historically been studied) emphasises the need to attract and empower functional biologists: can all 13 500 genes in Drosophila really be explained in terms of developmental biology? So,by embracing the integrative physiology manifesto, comparative physiologists can not only accelerate their own research, but their functional skills can make them indispensable in the post-genomic endeavour.

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Cloning, Characterization, and Expression of a G-Protein-Coupled Receptor fromLymnaea stagnalisand Identification of a Leucokinin-Like Peptide, PSFHSWSamide, as Its Endogenous Ligand

Cited by 73 publications

References 41 publications

Functional characterisation of theAnophelesleucokinins and their cognate G-protein coupled receptor

Functional characterisation of theAnophelesleucokinins and their cognate G-protein coupled receptor

Systematic G-protein-coupled Receptor Analysis inDrosophila melanogaster Identifies a Leucokinin Receptor with Novel Roles

Integrative physiology, functional genomics and the phenotype gap: a guide for comparative physiologists

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