Analysis of cardiovascular development and pathophysiology by gene engineering of the endothelin system in mice

通过内皮素系统基因工程分析小鼠心血管发育和病理生理学

基本信息

批准号：
18390229
负责人：
KURIHARA Hiroki
金额：
$ 11.36万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

We have achieved the following results in regard to the molecular mechanisms underlying the involvement of the endothelin (ET) system in cardiovascular and craniofacial development and pathophysiology.1. We have established the Cre-recombinase-mediated gene knock-in system in mouse ES cells, in which we can systematically replace the ET-A receptor (STAR) gene with exogenous genes.2. By using this system, we knocked-in cDNAs encoding the ET-B receptor (ETBR) and ETAR-ETBR chimeric receptors. These experiments have revealed that i) both STAR subtype-selective and nonselective signaling are involved in the craniofacial development and ii) the induction of Dlx5/Dlx6 homeobox genes and subsequent specification of the mandibular identity is mediated by the STAR subtype-selective, Gq/G11-mediated signaling pathway.3. Knock-in of the lacZ gene revealed a possible novel cell lineage originating within the cardiac crescent and contributing to the early cardiovascular development.4. Knock-in of EGFP enabled us to visualize the STAR-positive cells in situ in mice, which is useful for the analysis of the dynamics of STAR-positive cells (e.g. smooth muscle cells) in embryonic vascular formation and in the physiological and pathophysiological processes.5. We have identified Calpain-6 as a target molecule of the ET-1/STAR signaling pathway in craniofacial development. We have discovered the novel function of Calpain-6 in the stabilization of microtubules and actin organization involved in cellular morphology and motility. The studies on the developmental role of Calpain-6 are starting with its knockout mice established recently.These achievements contribute to the understanding of the mechanisms of the cardiovascular and craniofacial development and pave a way to the development of experimental systems applicable to the studies on (patho)physiology involving the ET system.

关于内皮素（ET）系统参与心血管和颅面发育和病理生理学的分子机制，我们已经取得了以下结果。1。我们已经在小鼠ES细胞中建立了CRE成年酶介导的基因敲入系统，在该系统中我们可以系统地用外源基因替代ET-A受体（Star）基因2。通过使用该系统，我们敲打编码ET-B受体（ETBR）和ETAR-ETBR嵌合受体的cDNA。这些实验表明，i）i）恒星亚型选择性和非选择性信号与颅面发育有关，ii）诱导DLX5/dlx6同源蛋白蛋白蛋白蛋白蛋白酶基因以及随后的下颌身份的规范，由星形亚型，GQ/类型，GQ/TYPE，GQ/Typeception介导G11介导的信号通路3。 LACZ基因的敲入显示出可能起源于心脏新月内的新细胞谱系，并导致早期心血管发育。4。 EGFP的敲击使我们能够可视化小鼠的恒星阳性细胞原位，这对于分析胚胎血管形成和生理和病理生理过程中星形阳性细胞（例如平滑肌细胞）的动力学非常有用.5。我们已经确定Calpain-6是颅面发育中ET-1/Star信号通路的靶分子。我们发现了Calpain-6在微管和肌动蛋白组织稳定中的新功能，参与细胞形态和运动。关于Calpain-6的发育作用的研究是从最近建立的敲除小鼠开始。这些成就有助于理解心血管和颅面发育的机制，并为适用于研究的实验系统开发的方式铺平了研究（（病情）涉及ET系统的生理学。