Molecular cloning and pharmacological characterization of the guinea pig 5-HT1E receptor
Molecular cloning and pharmacological characterization of the guinea pig 5-HT1E receptor
复制标题
DOI：
10.1016/j.ejphar.2003.11.019
复制
发表时间：
2004-01-26
影响因子：
5
通讯作者：
Nelson, DL
中科院分区：
文献类型：
--
作者：
Bai, F;Yin, TG;Nelson, DL
人类5-HT1E受体基因是十多年前克隆的。人们对它的功能知之甚少，也没有关于它存在于小型实验室动物基因组中的报道。在这项研究中，试图从大鼠和小鼠身上克隆5-HT1E基因的尝试没有成功。事实上，对小鼠基因组数据库的搜索显示，5-HT1E受体基因在小鼠基因组中缺失。然而，从豚鼠基因组DNA中克隆了5-HT1E基因，并对其进行了鉴定。豚鼠5-HT1E受体基因编码365个氨基酸的蛋白质。它与人类受体有88%(核酸)和95%(氨基酸)的同源性。在放射配基结合实验中，豚鼠5-HT1E受体显示出与人5-HT1E受体相似的药理作用。5-羟色胺(5-羟色胺，5-HT)剂量依赖性地刺激[S-35]GTP与豚鼠5-HT1E受体结合，其EC50为13.6+/-1.92 nM，与人5-HT1E受体的EC50(13.7+/-1.78 nM)相似。麦角新碱、α-甲基-5-羟色胺、1-萘基哌嗪、甲基丝氨酸、色胺和1-(2，5-二甲氧基-4-碘苯基)-2-氨基丙烷(DOI)也可激活豚鼠5-HT1E受体。甲硫西平具有拮抗活性。实时定量聚合酶链式反应(qRT-PCR)分析表明，5-HT1E基因在豚鼠大脑中的表达丰度最高的是海马区，其次是嗅球。在皮质、丘脑、脑桥、下丘脑、中脑、纹状体和小脑中检测到较低的水平。我们目前的研究标志着在常用的实验动物物种中首次鉴定出5-HT1E受体基因。这一发现应该允许阐明受体在中枢5-羟色胺能效应的复杂协调中的作用(S)。(C)2003爱思唯尔B.V.保留所有权利。
The human 5-HT1E receptor gene was cloned more than a decade ago. Little is known about its function, and there have been no reports of its existence in the genome of small laboratory animals. In this study, attempts to clone the 5-HT1E gene from the rat and mouse were unsuccessful. In fact, a search of the mouse genome database revealed that the 5-HT1E receptor gene is missing from the mouse genome. However, the 5-HT1E gene was cloned from guinea pig genomic DNA and was characterized. The guinea pig 5-HT1E receptor gene encodes a protein of 365 amino acids. It shares 88% (nucleic acid) and 95% (amino acid) homology with the human receptor. The guinea pig 5-HT1E receptor showed similar pharmacology to the human 5-HT1E receptor in radioligand binding assays. Serotonin (5-hydroxytryptamine, 5-HT) dose-dependently stimulated [S-35]GTPgammaS binding to the guinea pig 5-HT1E receptor with an EC50 of 13.6 +/- 1.92 nM, similar to that of the human 5-HT1E receptor (13.7 +/- 1.78 nM). Activation of the guinea pig 5-HT1E receptor was also achieved by ergonovine, alpha-methyl-5-HT, 1-naphthylpiperazine, methysergide, tryptamine, and 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI). Methiothepin exhibited antagonist activity. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that 5-HT1E mRNA was present in the guinea pig brain with the greatest abundance in the hippocampus, followed by the olfactory bulb. Lower levels were detected in the cortex, thalamus, pons, hypothalamus, midbrain, striatum, and cerebellum. Our current study marks the first identification of the 5-HT1E receptor gene in a commonly used laboratory animal species. This finding should allow the elucidation of the receptor's role(s) in the complex coordination of central serotonergic effects. (C) 2003 Elsevier B.V All rights reserved.