Gene targeting of natriuretic peptide family and their functional analyzes

利尿钠肽家族的基因打靶及其功能分析

• 批准号: 05680569
• 负责人: MINAMINO Naoto
• 金额: $ 1.28万
• 依托单位: National Cardiovascular Center Research Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05680569/
• 关键词: Natriuretic peptide; Gene targeting; Natriuretic peptide receptor; Gene structure analysis; Blood pressure regulation; Growth and differentiation; Molecular Biology; 機能解析; 遺伝子構造; マウス

Natriuretic peptide family is comprised of three peptides (ANP,BNP,CNP). These peptides bind receptor quanylate cyclase (GC-A and GC-B), increase cGMP concentration, and alter cell functions. ANP and BNP mainly function as cardiac hormones regulating blood pressure, while CNP is a neuropeptide in brain or a local regulator in the vascular wall. In order to elucidate their in vivo functions and estimate the contribution to blood pressure regulation, we have planned to establish a mouse which is deleted the natriuretic peptide genes by using the newly developed gene targeting method.We cloned mouse ANP,BNP and CNP gene of 5-6 kb containing complete sets of each exons. About 3-6 kb sequence of each gene was determine. In the case of CNP gene, the third exon region was first sequenced by the present study. In addition to three natriuretic peptide genes, we further cloned mouse GC-A and GC-B gene to analyze their functions. Portions of GC-A and GC-B gene structure containing functional domains are now being determined. By a series of these studies, appropriate fragments used for the gene targeting of the natriuretic peptides and their receptors have been obtained, and most of the sequences have already determined.

利钠肽家族由三种肽（ANP、BNP、CNP）组成。这些肽结合受体环化酶（GC-A和GC-B），增加cGMP浓度，改变细胞功能。ANP和BNP主要作为心脏激素调节血压，而CNP是脑内的神经肽或血管壁的局部调节剂。为了阐明它们在体内的功能，估计它们对血压调节的贡献，我们计划用新开发的基因靶向方法建立一个删除利钠肽基因的小鼠。我们克隆了5-6 kb的小鼠ANP、BNP和CNP基因，每个基因都包含完整的外显子。每个基因的序列约为3-6 kb。以CNP基因为例，本研究首先对其第三外显子区域进行了测序。除了三个利钠肽基因外，我们还克隆了小鼠GC-A和GC-B基因来分析它们的功能。GC-A和GC-B基因结构中包含功能域的部分正在被确定。通过一系列的研究，获得了用于利钠肽及其受体基因靶向的合适片段，并确定了大部分序列。

项目成果

M.Kojima et al.: "Cloning and characterization of a novel natriuretic peptide in frog (Rana catesbeiana)." J.Biol.Chem.269. 13136-13140 (1994)

M.Kojima 等人：“青蛙 (Rana catesbeiana) 中新型利尿钠肽的克隆和表征。”

S.Sugo et al.: "Endothelial cells actively synthesize and secrete adrenomedullin." Biochem.Biophys.Res Commun.201. 1160-1166 (1994)

S.Sugo 等人：“内皮细胞主动合成和分泌肾上腺髓质素。”

S.Sugo, et al.: "Production and secretion of adrenomedullin from vascular smooth muscle cells : Augmented production by tumor necrosis factor-alpha." Biochem.Biophys.Res.Commun.203. 719-726 (1994)

S.Sugo 等人：“血管平滑肌细胞产生和分泌肾上腺髓质素：通过肿瘤坏死因子-α 增强肾上腺髓质素的产生。”

T.Nishikimi,et al.: "Role of endogenous strial natriuretic peptide on systemic and renal hemodynamics in heart failure rats;Endogenous ANP in heart failu" Am.J.Physiol.267. H182-H186 (1994)

T.Nishikimi 等人：“内源性心房钠尿肽对心力衰竭大鼠全身和肾脏血流动力学的作用；心力衰竭中的内源性 ANP”Am.J.Physiol.267。

S.Sugo et al.: "Production and secretion of adrenomedullin from vascular smooth muscle cells:Augmented production by tumor necrosis factor-α" Biochem.Biophys.Res.Commun.203. 719-726 (1994)

S.Sugo 等人：“血管平滑肌细胞肾上腺髓质素的产生和分泌：通过肿瘤坏死因子-α 增强产生”Biochem.Biophys.Res.Commun.203 (1994)。