Bach A W, Unger L, Sprengel R, Mengod G, Palacios J, Seeburg P H, Voigt M M
BASF AG, Dept. Biotechnology ZHB, Ludwigshafen, FRG.
J Recept Res. 1993;13(1-4):479-502. doi: 10.3109/10799899309073674.
Using polymerase chain reaction (PCR) a complementary DNA (cDNA) encoding a 5-hydroxytryptamine (5-HT) receptor was isolated from rat forebrain. The amplified cDNA specifies an open reading frame of 374 amino acids comprising seven putative transmembrane regions. Expression of the cloned cDNA in human embryonic kidney cells (HEK 293) was used to establish the pharmacological profile of the encoded receptor polypeptide. Membranes containing the cloned receptor showed high affinity binding of [3H]-5-HT. Competition binding experiments with a variety of serotonin receptor ligands displayed a rank order of affinities corresponding to a 5-HT1D subtype: 5-CT > 5-HT = metergoline > CGS 12066 > methysergide > sumatriptan > mianserin = (-)alpha-Me-5-HT = yohimbine > 8-OH-DPAT > or = rauwolscine > spiperone > DOI > propranolol > or = 2-Me-5-HT > or = ICS 205930. Ketanserin and ritanserin displaced [3H]-5-HT-binding in a biphasic manner. In situ hybridization revealed highest expression of the corresponding mRNA in the pyramidal layer of the olfactory tubercle and the nucleus caudatus and accumbens.
利用聚合酶链反应（PCR）从大鼠前脑分离出编码5-羟色胺（5-HT）受体的互补DNA（cDNA）。扩增得到的cDNA确定了一个由374个氨基酸组成的开放阅读框，包含七个假定的跨膜区域。通过在人胚肾细胞（HEK 293）中表达克隆的cDNA，来确定所编码的受体多肽的药理学特征。含有克隆受体的细胞膜显示出对[3H]-5-HT的高亲和力结合。用多种5-羟色胺受体配体进行的竞争结合实验显示出的亲和力排序对应于5-HT1D亚型：5-CT > 5-HT = 美替拉酮 > CGS 12066 > 麦角新碱 > 舒马曲坦 > 米安色林 = (-)α-甲基-5-HT = 育亨宾 > 8-羟基二丙胺 > 或 = 萝芙木碱 > 螺哌隆 > DOI > 普萘洛尔 > 或 = 2-甲基-5-HT > 或 = ICS 205930。酮色林和利坦色林以双相方式取代[3H]-5-HT结合。原位杂交显示相应mRNA在嗅结节锥体细胞层、尾状核和伏隔核中表达最高。
