Molecular cloning and functional expression of the Penaeus monodon 5-HT receptor

Chalermporn Ongvarrasopone , Yaowaluck Roshorm , Suthasinee Somyong , Chetsada Pothiratana , Soontaree Petchdee , Jarasporn Tangkhabuanbutra , Samaisukh Sophasan , Sakol Panyim
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引用次数: 41

Abstract

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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单对虾5-HT受体的分子克隆及功能表达
5-羟色胺(5-HT)通过与各种5-HT受体亚型相互作用,介导甲壳类动物的多种生理功能。从黑虎对虾(Penaeus monodon)卵巢中克隆出一个5-羟色胺受体,包含2291个核苷酸,编码591个氨基酸的5-羟色胺受体蛋白。5-HT1Pem在HEK293细胞中瞬时表达,显示了饱和[3H]-5-HT结合,Kd为10.43±1.13 nM, Bmax为1.53±0.06 pmol/mg。假设的5-HT1Pem受体在所有被检查的组织中表达,并在卵巢成熟和成熟期组成性表达。针对5-HT受体第三胞内环(i3环)的多克隆抗体显示,5-HT1Pem受体蛋白在卵巢1期和2期的小梁中表达,而在卵巢3期和4期的皮质杆和卵母膜周围表达,表明受体定位在调节对虾卵巢成熟和产卵中起关键作用。
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