Identification of histamine H_1 receptor subtypes

组胺H_1受体亚型的鉴定

• 批准号: 07557194
• 负责人: FUKUI Hiroyuki
• 金额: $ 1.66万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557194/
• 关键词: histamine; receptor; adrenal medulla; cloning; Xenopus Laevis oocyte; ヒスタミンH_1受容体; 受容体サブタイプ

mRNA derived from bovine adrenal medulla was fractionated by the sucrose density gradient centrifugation, and each fraction was injected into Xenopus Laevis oocytes. Expression of histamine H_1 receptors was electrophysiologically examined. Injection of fractions containing 3.5-kb mRNA or 5.0-kb mRNA induced the expression of histamine H_1 receptors. This result suggests the existence of histamine H_1 receptor subtypes. Histamine H_1 receptor-dependent inward Cl^- current was aprox. 50nA when 3.5-kb mRNA was injected, and smaller than 40nA when 5.0-kb mRNA was injected. cDNA libraries derived from 3.5-kb mRMA and 5.0-kb mRNA were constructed using lambda ZAPII,a vector for in vitro transcription. The cDNA library was screened by the injection of mRNA through in vitro transcription and electrophysiological detection of histamine H_1 receptor expression. The histamine H_1 receptor cDNA derived from 3.5-kb mRNA was isolated, and the structure of the receptor was determined. The detection of histamine H_1 receptor-dependent inward Cl^- current from 5.0-kb mRNA fraction was not succeeded when mRNA through in vitro transcription was injected. Recently histamine H_1 receptor knock out mouse was prepared, and the lack of histamine H_1 receptors was demonstrated by the autoradiography of [^3H] mepyramine binding. This result indicated that mouse has only one histamine H_1 receptor gene. We observed that human and guinea pig have two kinds of histamine H_1 rceptor mRNAs with different molecular weights. Physiological and pathological roles of histamine differ from species, and the possibility of histamine H_1 receptor subtypes in human and guinea pig still remains.

采用蔗糖密度梯度离心法分离牛肾上腺髓质的mRNA，并将各组分注射到非洲爪蟾卵母细胞中。电生理检测组胺H_1受体表达。注射含3.5kb或5.0kb mRNA的组分可诱导组胺H_1受体的表达。提示组胺H_1受体亚型的存在。组胺H_1受体依赖性内向Cl^-电流接近。当注射3.5-kbmRNA时，50 nA小于40 nA，当注射5.0-kbmRNA时，50 nA小于40 nA。使用体外转录载体Lambda ZAPII构建了源自3.5-kb mRMA和5.0-kb mRNA的cDNA文库。通过体外转录和组胺H_1受体表达的电生理检测，注射mRNA筛选cDNA文库。从3.5kb的mRNA中分离出组胺H_1受体cDNA，并测定了受体的结构。当注射经体外转录的mRNA时，从5.0 kb的mRNA片段中检测到组胺H_1受体依赖的内向Cl^-电流并不成功。近年来，我们成功地制备了组胺H_1受体基因敲除小鼠，并通过[^3H]美托咪胺结合放射自显影证实了小鼠体内组胺H_1受体的缺失。这一结果表明小鼠只有一个组胺H_1受体基因。我们观察到人和豚鼠有两种分子量不同的组胺H_1受体mRNA。组胺的生理和病理作用因物种而异，人和豚鼠存在组胺H_1受体亚型的可能性仍然存在。

项目成果

渡辺建彦: "ヒスタミン受容体、蛋白質核酸酵素" 共立出版, 8 (1997)

渡边立彦：“组胺受体、蛋白质核酸酶”共立出版，8（1997）

Takagishi,T: "Expression of the histamine H_1 receptor gene in relation to athelosclerosis." Am.J.Pathol.146. 981-988 (1995)

Takagishi,T：“与动脉粥样硬化相关的组胺 H_1 受体基因的表达。”

Takagishi, T,Sasaguri, Y,Nakano, R,Arima, N,Tanimoto, A,Fukui, H and Morimatsu, M: "Expression of the histamine H_1 receptor gene in relation to athelosclerosis" Am J Pathol. 146. 981-988 (1995)

Takagishi，T，Sasaguri，Y，Nakano，R，Arima，N，Tanimoto，A，Fukui，H 和 Morimatsu，M：“与动脉粥样硬化相关的组胺 H_1 受体基因的表达”Am J Pathol。

Nonaka, H,Otaki, S,Ohshima, E,Kono, M,Kase, H,Ohta, K,Fukui, H and Ichimura, M: "Unique binding pocket for KW-4679 in the histamine H_1 receptor" Eur J Pharmacol. (in press).

Nonaka, H,Otaki, S,Ohshima, E,Kono, M,Kase, H,Ohta, K,Fukui, H 和 Ichimura, M：“组胺 H_1 受体中 KW-4679 的独特结合口袋”Eur J Pharmacol。

Nonaka,H.: "Unique binding pocket for KW-4679 in the histamine H^1 receptor" Eur.J.Pharmacol.(in press).

Nonaka, H.：“组胺 H^1 受体中 KW-4679 的独特结合袋”Eur.J.Pharmacol.（出版中）。