title: Characterization of a tyramine receptor type 2 from hemocytes of rice stem borer, Chilo suppressalis.
abstract: Calcium acts as a second messenger in many cell types, including insect hemocytes. Intracellular calcium level has a definite role in innate and adaptive immune signaling. Biogenic amines such as octopamine (OA), tyramine (TA), dopamine (DA) and serotonin (5-HT) play various important physiological roles in insects by activating distinct G-protein-coupled receptors (GPCRs) that share a putative seven transmembrane domain structure. OA and 5-HT have been shown that can mediate insect hemocytic immune reactions to infections and invasions. Here, we showed that TA increase hemocyte spreading in the rice stem borer, Chilo suppressalis. Furthermore, we cloned a cDNA encoding a tyramine receptor type 2 from the hemocytes in the C. suppressalis, viz., CsTA2, which shares high sequence similarity to members of the invertebrate tyramine receptor family. The CsTA2 receptor was stably expressed in human embryonic kidney (HEK) 293 cells, and its ligand response has been examined. Receptor activation with TA induced a dose-dependent increase in intracellular Ca(2+) concentration ([Ca(2+)]i) in cells, with an EC50 value of 18.7±5.3 nM, whereas OA, DA, 5-HT and other potential agonists did not have this response. The mRNA is present in various tissues including nerve cord, hemocytes, fat body, midgut, Malpighian tubules, and epidermis in the larval stage. Western blot analysis and immunohistochemistry assay displayed that CsTA2 was detected and presented on hemocytes. We also showed that TA induced Ca(2+) release from the hemocytes of C. suppressalis.
authors.affiliation: State & Local Joint Engineering Research Center of Green Pesticide Invention and Application, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; State Key Laboratory of Rice Biology & Key Laboratory of Agricultural Entomology of Ministry of Agriculture, Institute of Insect Sciences, Zhejiang University, Hangzhou, China. Electronic address: wusf@njau.edu.cn.
authors.forename: Shun-Fan
authors.initials: SF
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Rice Biology & Key Laboratory of Agricultural Entomology of Ministry of Agriculture, Institute of Insect Sciences, Zhejiang University, Hangzhou, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Rice Biology & Key Laboratory of Agricultural Entomology of Ministry of Agriculture, Institute of Insect Sciences, Zhejiang University, Hangzhou, China. Electronic address: chu@zju.edu.cn.
authors.forename: Gong-Yin
authors.initials: GY
authors.lastname: Ye
mesh_terms: D000818:Animals; D002118:Calcium / Q000502:physiology; D018076:DNA, Complementary / Q000235:genetics; D004298:Dopamine / Q000494:pharmacology; D057809:HEK293 Cells; D006434:Hemocytes / Q000378:metabolism*; D006801:Humans; D019476:Insect Proteins / Q000378:metabolism; D007814:Larva / Q000378:metabolism; D009036:Moths / Q000235:genetics / Q000378:metabolism*; D009655:Octopamine / Q000494:pharmacology; D017982:Receptors, Biogenic Amine / Q000235:genetics / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D015290:Second Messenger Systems; D012701:Serotonin / Q000494:pharmacology; D014439:Tyramine / Q000494:pharmacology*
chemical_list: D018076:DNA, Complementary; D019476:Insect Proteins; D017982:Receptors, Biogenic Amine; D043562:Receptors, G-Protein-Coupled; C066175:tyramine receptor; D009655:Octopamine; D012701:Serotonin; D002118:Calcium; D004298:Dopamine; D014439:Tyramine
keywords: Chilo suppressalis; G protein-coupled receptor; Hemocytes; Octopamine; Tyramine