Characterization of a novel G-protein coupled receptor from the parasitic nematode H. contortus with high affinity for serotonin.
The neurotransmitter serotonin (5HT) has been shown to modulate mobility, feeding, egg-laying, and defecation behaviors in the saprophytic nematode Caenorhabditis elegans. Although the effects of serotonin on these behaviors in parasitic nematodes is under study, little is known about the diversity, ontogeny, signaling, and pharmacology of serotonin receptors in these organisms. In an effort to increase our understanding of this system, we cloned and characterized a novel cDNA (5HT1Hc) from the parasitic nematode Haemonchus contortus that has high amino acid sequence homology with known G-protein coupled 5HT1-receptors from invertebrates and vertebrates. Transcript expression studies in four development stages (egg, L1/L2, L3, and adult) revealed the presence of the mRNA in the L1/L2, L3, and adult stages. Membranes from insect cells (Sf9) expressing the 5HT1Hc-receptor cDNA displayed nanomolar binding affinity to serotonin and a unique pharmacological profile distinct from known invertebrate and mammalian 5HT-receptors. Receptor signaling studies with mammalian AV12 cells expressing the 5HT1Hc-receptor and the promiscuous G-protein, Galpha15, demonstrated dose-dependent intracellular signals with serotonin acting as an agonist. Together, these studies describe a novel invertebrate 5HT-receptor with high affinity for the indolealkylamine, serotonin, and pharmacological properties that do not conform to any known members of this superfamily of metabotropic receptors.
Smith
Martin W
MW
Eli Lilly and Company, Sphinx Laboratories, Research Triangle Park, North Carolina, USA.
Borts
Tracy L
TL
Emkey
Renee
R
Cook
Carolyn A
CA
Wiggins
Christina J
CJ
Gutierrez
Jesus A
JA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive / Q000502:physiology; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D019204:GTP-Binding Proteins / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D006189:Haemonchus / Q000235:genetics*; D008024:Ligands; D008969:Molecular Sequence Data; D010802:Phylogeny; D011985:Receptors, Serotonin / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D044263:Receptors, Serotonin, 5-HT1; D017422:Sequence Analysis, DNA; D012701:Serotonin / Q000737:chemistry / Q000378:metabolism*
D018076:DNA, Complementary; D008024:Ligands; D011985:Receptors, Serotonin; D044263:Receptors, Serotonin, 5-HT1; D012701:Serotonin; D019204:GTP-Binding Proteins