A W Bach, L Unger, R Sprengel, G Mengod, J Palacios, P H Seeburg, M M Voigt
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引用次数: 41
摘要
采用聚合酶链反应(PCR)技术从大鼠前脑中分离到编码5-羟色胺(5-HT)受体的互补DNA (cDNA)。扩增的cDNA指定一个374个氨基酸的开放阅读框，包括7个假定的跨膜区域。利用克隆的cDNA在人胚胎肾细胞(HEK 293)中的表达，建立编码受体多肽的药理学谱。含有克隆受体的膜与[3H]-5-HT具有高亲和力。与多种5-羟色胺受体配体的竞争结合实验显示，5-羟色胺受体配体的亲和顺序为:5-CT > 5-羟色胺=美高林> CGS 12066 >甲基塞吉胺>舒马匹坦>米安色林= (-)α - me -5-羟色胺=育亨宾> 8-OH-DPAT >或=毛wolsciine > spiperone > DOI >普萘洛尔>或= 2- me -5-羟色胺>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在嗅结节锥体层、尾状核和伏隔核中表达量最高。
Structure functional expression and spatial distribution of a cloned cDNA encoding a rat 5-HT1D-like receptor.
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.
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