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Molecular Anatomy of Functional Differentination Affected by Combination of Gap Junctions, Connexins

间隙连接、连接蛋白组合影响功能分化的分子解剖学

基本信息

批准号：
15390057
负责人：
SHIBATA Yosaburo
金额：
$ 9.73万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15390057/
关键词：
gene targeting heart gap junction connexin Cx45 Nfatcl cushion defect cardiac development コネキシンギャップ結合心臓 cardiac valve shaer stress heart

项目摘要

Cx45-deficient ES cells were induced to differentiate into contracting cardiac myocytes in vitro. Their contractions were uncoordinated due to defective intercellular communication. In vivo, we created mice lacking Cx45 in all the somatic cells (Cx45-KO), in cardiac myocytes (Cx45-CA), and in vascular endothelial cells (Cx45-Tie2). We found intriguing differences of phenotypes in each mutant. The Cx45-KO mice died at the E10 stage, showing atrioventricular conduction block and an endocardial cushion defect in the early heart. The Ca2+-dependent transcription factor Nfatcl was in inactive, cytoplasmic form in the Cx45-KO endocardial endothelium. The Cx45-CA mice died at the E10 stage, showing atrioventricular conduction block, but without the endocardial cushion defect. Nfatcl was in active nuclear form in the Cx45-CA endocardial endothelium. The lethal stage shows that the conduction block is the primary cause of death in the Cx45-KO mice. The Cx45-Tie2 mice, in contrast, did not show … More any abnormality. Moreover, we created mice lacking another cardiac connexin Cx43, in the absence of Cx45. They were identical to the Cx45-KO mice, indicating that Cx45 constitutes the most significant gap junction protein in the early heart.Next, we created cardiac troponin T (cTnT)-deficient mice. Because it is an essential component of the contracting apparatus, the cTnT-deficient mice did not show any contractions. Their lethal stage was the same as the above Cx45-mutants. They showed the endocardial cushion defect due to retarded cardiac development. Nfatcl localization, however, indicated active nuclear form.The endocardial cushion defect was only apparent when Cx45 was absent in both of the myocardial and the endocardial layer. Our model, in which Cx45 regulates the epithelial-mesenchymal transformation in the endocardial endothelium, needs further revise by the future study, though the defect is not caused by growth retardation. Collaborations with the overseas coworkers are in progress, revealing the tissue specific functions of Cx45 in neuronal/glial/neural crest cells. Roles of Cx45 in these cell types will be evident in the near future. Less

Cx45缺陷的ES细胞在体外被诱导分化为收缩的心肌细胞。由于细胞间通讯有缺陷，它们的收缩不协调。在体内，我们创建了所有体细胞（Cx45-KO）、心肌细胞（Cx45-CA）和血管内皮细胞（Cx45-Tie2）中缺乏Cx45的小鼠。我们发现每个突变体的表型存在有趣的差异。 Cx45-KO小鼠在E10阶段死亡，表现出早期心脏的房室传导阻滞和心内膜垫缺损。 Ca2+依赖性转录因子Nfatcl在Cx45-KO心内膜内皮中处于失活的细胞质形式。 Cx45-CA小鼠在E10期死亡，显示房室传导阻滞，但无心内膜垫缺损。 Nfatcl 在 Cx45-CA 心内膜内皮中呈活跃核形式。致死阶段表明传导阻滞是Cx45-KO小鼠死亡的主要原因。相比之下，Cx45-Tie2 小鼠没有表现出任何异常。此外，我们在缺乏 Cx45 的情况下培育了缺乏另一种心脏连接蛋白 Cx43 的小鼠。它们与 Cx45-KO 小鼠相同，表明 Cx45 构成早期心脏中最重要的间隙连接蛋白。接下来，我们创建了心肌肌钙蛋白 T (cTnT) 缺陷小鼠。因为它是收缩装置的重要组成部分，所以 cTnT 缺陷的小鼠没有表现出任何收缩。它们的致死阶段与上述Cx45突变体相同。他们显示出由于心脏发育迟缓而导致的心内膜垫缺陷。然而，Nfatcl 定位表明活跃的核形式。仅当心肌层和心内膜层均不存在 Cx45 时，心内膜垫缺陷才明显。我们的模型中，Cx45 调节心内膜内皮细胞的上皮间质转化，尽管该缺陷不是由生长迟缓引起的，但仍需要未来的研究进一步修改。与海外同事的合作正在进行中，揭示了Cx45在神经元/神经胶质/神经嵴细胞中的组织特异性功能。 Cx45 在这些细胞类型中的作用将在不久的将来变得明显。较少的

项目成果

期刊论文数量（28）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

M.Hirata et al.: "Pigment cell organization in the hypodermis of zebrafish"Dev.Dyn.. 227. 497-503 (2003)

M.Hirata 等：“斑马鱼皮下组织中的色素细胞组织”Dev.Dyn.. 227. 497-503 (2003)

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Shear-stress induced up-regulation of connexin 43 expresson in endothelial cells on upstream surfaces of rat cardiac valves.

剪切应力诱导大鼠心脏瓣膜上游表面内皮细胞连接蛋白 43 表达上调。

DOI：
发表时间：
2004
期刊：
Histochem.Cell Biol. 122
影响因子：
0
作者：
T.Inai;M.R.Mancuso;D.M.McDonald;J.Kobayashi;K.Nakamura;Y.Shibata;S.Mori;Naoki Ishiguro. et al.;T.Inai et al.
通讯作者：
T.Inai et al.

K.Nishii et al.: "Mice lacking connexin45 conditionally in cardiac myocytes display embryonic lethality similar to that of germline knockout mice without endocardial cushion defect"Cell Commun.Adhesion. 10. 365-369 (2003)

K.Nishii 等人：“心肌细胞条件性缺乏连接蛋白 45 的小鼠表现出与没有心内膜垫缺陷的种系基因敲除小鼠相似的胚胎致死率”Cell Commun.Adhesion。