Cloning and functional analysis of the mouse vasopressin/angiotensin II dual receptor

小鼠加压素/血管紧张素II双受体的克隆及功能分析

• 批准号: 17590964
• 负责人: IWASAKI Yasumasa
• 金额: $ 2.24万
• 依托单位: Kochi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590964/
• 关键词: vasopressin; angiotensin; receptor; NLR receptor; LRR protein; NACHT; アンギオテンシン; レニン; シグナル伝達

Angiotensin II/vasopressin dual receptor (AT/VPR) gene was cloned from the rat, the protein product of which has unique receptor characteristics, i.e. both angiontensin II (AII) and vasopressin (VP) act as ligands. However, the structure of the gene has not characterized yet, and the functional characteristics of the receptor remains to be clarified. In this study, we cloned the mouse AT/VPR mRNA and gene (6.5 Kb). Although the mouse and the rat AT/AVPR genes showed extremely high similarity in their nucleotide sequences, unexpectedly, the initiation codon of the rat gene was changed from Met to Leu in the mouse, resulting in extended protein product at N-terminus (179 a.a.). The mouse AT/VPR gene consisted of at least 5 exons and 4 introns by comparing the nucleotide sequence of its cDNA. Interestingly, the gene partly shared that of NACHT/Nalp6,which belongs the pathogen-recognition receptor.We then examined the tissue distribution of the AT/VPR mRNA, and found it to be expressed in the pituitary, thalamus, cerebral cortex, hippocampus, cerebellum, liver, gall bladder, adrenal gland, colon, testis, and white adipose tissue.Expression in cardiomyocytes was weak, and no expression was observed in skeletal muscle, skin, and spleen.Finally, we constructed an expression vector of the receptor and expressed it in A10 rat vascular smooth muscle cells in vitro. We fund that, at least in our experimental condition, AII did not activate any intracellular signaling pathways. On the other hand, vasopressin modestly enhanced the response of SRE-dependent transcription, whereas markedly attenuated that of NF-kB-dependent transcription, when the AT/VPR was overexpressed.We thus conclude that AT/VPR may be working not as a classical "hormone receptor" but as a pattern recognition receptor, and is modifying the NF-kB-dependent gene transcription. The functional differences between the mouse and rat AT/VPR awaits further investigation.

血管紧张素II/加压素双重受体（AT/VPR）基因是从大鼠中克隆的，其蛋白产物具有独特的受体特性，即血管紧张素II（AII）和加压素（VP）均作为配体发挥作用。然而，该基因的结构尚未表征，受体的功能特征仍有待澄清。本研究克隆了小鼠AT/VPR的mRNA和基因（6.5Kb）。尽管小鼠和大鼠AT/AVPR基因在它们的核苷酸序列中显示出极高的相似性，但出乎意料的是，大鼠基因的起始密码子在小鼠中从Met变为Leu，导致N-末端的延伸蛋白产物（179 a.a.）。通过比较小鼠AT/VPR基因cDNA的核苷酸序列，发现该基因至少由5个外显子和4个内含子组成。有趣的是，该基因部分与病原体识别受体NACHT/Nalp 6的基因相同。我们随后检查了AT/VPR mRNA的组织分布，发现其在垂体、丘脑、大脑皮质、海马、小脑、肝脏、胆囊、肾上腺、结肠、睾丸和白色脂肪组织中表达。在心肌细胞中表达较弱，在骨骼肌中没有观察到表达。最后，我们构建了该受体的表达载体，并在体外培养的A10大鼠血管平滑肌细胞中进行了表达。我们发现，至少在我们的实验条件下，AII没有激活任何细胞内信号通路。另一方面，当AT/VPR过表达时，加压素适度增强SRE依赖的转录反应，而显著减弱NF-κ B依赖的转录反应，因此我们认为AT/VPR可能不是作为一种经典的“激素受体”，而是作为一种模式识别受体，并修饰NF-κ B依赖的基因转录。小鼠和大鼠AT/VPR的功能差异有待进一步研究。

项目成果

マウスバゾプレシン/アンジオテンシンデュアル受容体のクローニングと機能解析

小鼠加压素/血管紧张素双受体克隆及功能分析

• 发表时间: 2007
• 期刊: 日本内分泌学会雑誌 第80回日本内分泌学会学術総会抄録集 Vol 83,No 1(In press)
• 作者: 岩崎泰正;西山充;次田誠;谷口義典;岡崎瑞穂;何静;橋本浩三
• 通讯作者: 橋本浩三

マウスァシジォテンシン・バソプレシンデュアル受容体のクローニングと機能解析

小鼠担负压素/加压素双受体的克隆及功能分析

• 发表时间: 2007
• 期刊: 日本内分泌学会雑誌 第80回日本内分泌学会学術総会抄録集 Vol.83, No.1(In press)
• 作者: 岩崎泰正;西山充;次田誠;谷口義典;岡崎瑞穂;何静;橋本浩三
• 通讯作者: 橋本浩三

Folia Endocrinologia Japonica

日本叶内分泌学

• 发表时间: 2007
• 期刊: 80^<th> Japanese Endocrine Society Vol. 83, No. 1 (in press)
• 作者: Yasumasa Iwasaki;Mitsuru Nishiyama;Makoto Tsugita;Yoshinori Taniguchi;Mizuho Okazaki;He Jing;Kozo Hashimoto
• 通讯作者: Kozo Hashimoto