Cloning, functional expression, and mRNA tissue distribution of the rat 5-hydroxytryptamine1A receptor gene.
Cloning, functional expression, and mRNA tissue distribution of the rat 5-hydroxytryptamine1A receptor gene.
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DOI：
10.1016/s0021-9258(19)39437-2
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发表时间：
1990-04
期刊：
影响因子：
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通讯作者：
Paul R. Albert;Qun-Yong Zhou;H. V. Tol;J. Bunzow;O. Civelli
中科院分区：
文献类型：
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作者：
Paul R. Albert;Qun-Yong Zhou;H. V. Tol;J. Bunzow;O. Civelli
G蛋白偶联受体包括具有显著序列相似性的基因家族。我们已经筛选了大鼠基因组文库在低严格性杂交条件下与仓鼠β 2-肾上腺素能受体基因的编码部分，以分离该基因家族的新成员。其中克隆D编码5-HT 1A结合位点，因为：1）它具有一个无内含子的开放阅读框架，编码一个具有7个跨膜结构域的蛋白质，与人5-HT 1A受体（G21）的氨基酸同源性为89%; 2）当转染到Ltk-细胞中时，其表达具有5-HT 1A受体亚型的药理学的配体结合位点，包括8-羟基-5-HT-和螺唑嘌呤-可置换结合。（2-（N，N-二[2，3 - 3 H]丙基氨基）-1，2，3，4-四氢萘（KH = 0.8 nM）。我们进一步表明，克隆D编码一个功能性受体，因为它的结合位点与G蛋白相互作用，因为它介导激动剂诱导的抑制基础和刺激cAMP的积累在转染的GH4 C1垂体细胞。最后，我们分析了5-HT 1A受体mRNA在大鼠脑中的组织分布，发现5-HT 1A mRNA与5-HT 1A受体的预期分布（在隔区和海马中最高）一样存在，但以三种RNA种类（3.9、3.6和3.3种酶）存在。这些研究代表了克隆的大鼠5-HT 1A受体的受体功能和脑分布的第一个表征。
G protein-coupled receptors comprise a family of genes that share significant sequence similarity. We have screened a rat genomic library under low stringency hybridization conditions with the coding portion of the hamster beta 2-adrenergic receptor gene to isolate new members of this gene family. We show that one of these clones, clone D, codes for a 5-hydroxytryptamine1A (5-HT1A) binding site since: 1) it possesses an intronless open reading frame encoding a protein with seven putative transmembrane domains and 89% amino acid identity with the human 5-HT1A receptor (G21); 2) when transfected into Ltk- cells, it expresses a ligand-binding site with the pharmacology of the 5-HT1A receptor subtype, including 5-HT- and spiroxatrine-displaceable binding of 8-hydroxy-(2-(N,N-di[2,3-3H]propylamino)-1,2,3,4-tetrahydronaphthalene (KH = 0.8 nM). We further show that clone D encodes a functional receptor because its binding site interacts with G proteins and because it mediates agonist-induced inhibition of basal and stimulated cAMP accumulation in transfected GH4C1 pituitary cells. Finally, we have analyzed the tissue distribution of 5-HT1A receptor mRNA in rat brain and have found that 5-HT1A mRNA is present with the expected distribution of the 5-HT1A receptor (highest in septum and hippocampus) but is present as three RNA species (3.9, 3.6, and 3.3 kilobases). These studies represent the first characterization of receptor function and brain distribution of the cloned rat 5-HT1A receptor.