{"title":"单对虾5-HT受体的分子克隆及功能表达","authors":"Chalermporn Ongvarrasopone , Yaowaluck Roshorm , Suthasinee Somyong , Chetsada Pothiratana , Soontaree Petchdee , Jarasporn Tangkhabuanbutra , Samaisukh Sophasan , Sakol Panyim","doi":"10.1016/j.bbaexp.2006.07.004","DOIUrl":null,"url":null,"abstract":"<div><p>Serotonin (5-HT) mediates a number of diverse physiological functions in crustaceans by interacting with various 5-HT receptor subtypes. A putative 5-HT receptor cloned from the ovary of the black tiger prawn (<em>Penaeus monodon</em>) consisted of 2291 nucleotides, encoding a putative 5-HT<sub>1Pem</sub> receptor protein of 591 amino acids. Transient expression of 5-HT<sub>1Pem</sub> in HEK293 cells demonstrated a saturable [<sup>3</sup>H]-5-HT binding with a Kd of 10.43<!--> <!-->±<!--> <!-->1.13 nM and <em>B</em><sub>max</sub> of 1.53<!--> <!-->±<!--> <!-->0.06 pmol/mg. The putative 5-HT<sub>1Pem</sub> receptor is expressed in all tissues examined and is constitutively expressed in the ovary during ovarian maturation and spent phase. Polyclonal antibodies against the third intracellular loop (i3 loop) of the 5-HT receptor showed that the 5-HT<sub>1Pem</sub> receptor protein was expressed in the trabeculae of ovarian stages 1 and 2 but on the cortical rod and surrounding the oocyte membrane of stages 3 and 4, suggesting that receptor localization plays a critical role in regulating ovarian maturation and spawning in penaeus shrimp.</p></div>","PeriodicalId":100161,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression","volume":"1759 7","pages":"Pages 328-339"},"PeriodicalIF":0.0000,"publicationDate":"2006-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bbaexp.2006.07.004","citationCount":"41","resultStr":"{\"title\":\"Molecular cloning and functional expression of the Penaeus monodon 5-HT receptor\",\"authors\":\"Chalermporn Ongvarrasopone , Yaowaluck Roshorm , Suthasinee Somyong , Chetsada Pothiratana , Soontaree Petchdee , Jarasporn Tangkhabuanbutra , Samaisukh Sophasan , Sakol Panyim\",\"doi\":\"10.1016/j.bbaexp.2006.07.004\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Serotonin (5-HT) mediates a number of diverse physiological functions in crustaceans by interacting with various 5-HT receptor subtypes. A putative 5-HT receptor cloned from the ovary of the black tiger prawn (<em>Penaeus monodon</em>) consisted of 2291 nucleotides, encoding a putative 5-HT<sub>1Pem</sub> receptor protein of 591 amino acids. Transient expression of 5-HT<sub>1Pem</sub> in HEK293 cells demonstrated a saturable [<sup>3</sup>H]-5-HT binding with a Kd of 10.43<!--> <!-->±<!--> <!-->1.13 nM and <em>B</em><sub>max</sub> of 1.53<!--> <!-->±<!--> <!-->0.06 pmol/mg. The putative 5-HT<sub>1Pem</sub> receptor is expressed in all tissues examined and is constitutively expressed in the ovary during ovarian maturation and spent phase. Polyclonal antibodies against the third intracellular loop (i3 loop) of the 5-HT receptor showed that the 5-HT<sub>1Pem</sub> receptor protein was expressed in the trabeculae of ovarian stages 1 and 2 but on the cortical rod and surrounding the oocyte membrane of stages 3 and 4, suggesting that receptor localization plays a critical role in regulating ovarian maturation and spawning in penaeus shrimp.</p></div>\",\"PeriodicalId\":100161,\"journal\":{\"name\":\"Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression\",\"volume\":\"1759 7\",\"pages\":\"Pages 328-339\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2006-07-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/j.bbaexp.2006.07.004\",\"citationCount\":\"41\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0167478106001126\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0167478106001126","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Molecular cloning and functional expression of the Penaeus monodon 5-HT receptor
Serotonin (5-HT) mediates a number of diverse physiological functions in crustaceans by interacting with various 5-HT receptor subtypes. A putative 5-HT receptor cloned from the ovary of the black tiger prawn (Penaeus monodon) consisted of 2291 nucleotides, encoding a putative 5-HT1Pem receptor protein of 591 amino acids. Transient expression of 5-HT1Pem in HEK293 cells demonstrated a saturable [3H]-5-HT binding with a Kd of 10.43 ± 1.13 nM and Bmax of 1.53 ± 0.06 pmol/mg. The putative 5-HT1Pem receptor is expressed in all tissues examined and is constitutively expressed in the ovary during ovarian maturation and spent phase. Polyclonal antibodies against the third intracellular loop (i3 loop) of the 5-HT receptor showed that the 5-HT1Pem receptor protein was expressed in the trabeculae of ovarian stages 1 and 2 but on the cortical rod and surrounding the oocyte membrane of stages 3 and 4, suggesting that receptor localization plays a critical role in regulating ovarian maturation and spawning in penaeus shrimp.
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