ANALYSIS OF T-TUBULES AND INTERCALATED DISKS IN MYOCARDIUM OF VINCLULIN KO MICE

VINCLULIN KO小鼠心肌T型管和闰盘的分析

• 批准号: 7722416
• 负责人: ROBERT A ROSS
• 金额: $ 0.39万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7722416
• 关键词: Actins; Adhesions; Cardiac Myocytes; Cardiomyopathies; Cell membrane; Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; Cytoskeleton; Dilated Cardiomyopathy; Electron Microscopy; Embryo; Excision; Exons; Funding; Genes; Grant; Institution; Intercalated disc; Knock-out; Knockout Mice; Localized; Mediating; Muscle; Muscle Cells; Mutate; Myocardium; Patients; Protein Isoforms; Proteins; RNA Splicing; Research; Research Personnel; Resources; Sarcolemma; Source; Stress; Technology; United States National Institutes of Health; Variant; Ventricular; Vinculin; electron tomography; link protein; metavinculin; mouse model; myosin light chain 2; recombinase

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Electron microscopy and electron tomography is being utilized at NCMIR to analyze the structural abnormalities associated with vinculin KO mice. Vinculin is a ubiquitously expressed multiliganded protein that links the actin cytoskeleton to the cell membrane. Metavinculin (MVin), its muscle-specific splice variant isoform, is found to be mutated in patients with dilated cardiomyopathies. In myocytes, it is localized in protein complexes which anchor the contractile apparatus to the sarcolemma. Vinculin is found at intercalated discs (ICDs) as well as in cell-to matrix adhesions. Its function in the myocardium remains poorly understood. Therefore, we developed a mouse model with cardiac-myocyte-specific inactivation of the vinculin (Vcl) gene by using Cre-loxP technology. Since global knockout (KO) of vinculin is embryonic lethal and heterozygous global knockout of vinculin predispose to stress-induced cardiomyopathy, we generated cardiac-myocyte specific vinculin KO mice. Floxed Vin exon 3 excision was mediated by myosin light chain-2 ventricular (MLC2v) Cre recombinase expression.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 中心，不一定是研究者的机构。 NCMIR 使用电子显微镜和电子断层扫描来分析与纽蛋白 KO 小鼠相关的结构异常。纽蛋白是一种普遍表达的多配体蛋白，它将肌动蛋白细胞骨架与细胞膜连接起来。 Metavinculin (MVin) 是其肌肉特异性剪接变体亚型，被发现在扩张型心肌病患者中发生突变。在肌细胞中，它位于将收缩装置锚定到肌膜的蛋白质复合物中。纽蛋白存在于闰盘 (ICD) 以及细胞与基质的粘附中。 它在心肌中的功能仍然知之甚少。因此，我们利用 Cre-loxP 技术开发了一种心肌细胞特异性纽蛋白 (Vcl) 基因失活的小鼠模型。由于纽蛋白的整体敲除（KO）是胚胎致死性的，并且杂合的纽蛋白整体敲除易诱发应激性心肌病，因此我们产生了心肌细胞特异性纽蛋白KO小鼠。 Floxed Vin 外显子 3 切除是由肌球蛋白轻链 2 心室 (MLC2v) Cre 重组酶表达介导的。