CARDIAC PROGENITOR CELL DIFFERENTIATION

描述：(改编自《调查员摘要》)申请者希望 了解心源性转录因子在检测中的作用 和心肌细胞的发育。他已经鉴定并克隆了一个cdna。 编码一种心脏调节性转录因子，称为CLP-1，可能发挥作用 在心源性祖细胞发育过程中起着重要作用。这个 当前应用程序的既定目标是了解CLP-1在 通过寻址CLP-1在何时、何地和以何种身份执行其心脏发生 作为心脏生成的调节器。该提案有四个具体目标。 目标1旨在对结构和结构进行额外表征 CLP-1的功能特性及其与其潜在的生心性的关系 监管属性。申请人和他的同事将尝试 确定负责结合B的CLP-1的结构域 检测心脏MLC-2基因启动子中的MEF-2元件结合部位 这些相同的结构域介导了心脏MLC-2的CLP-1激活 基因在体内。目标2旨在研究细胞功能 CLP-1，以确定其作为心脏基因和 心肌细胞分化。这些研究将集中于确定1) CLP-1作为心源性转录激活剂的潜力 能够参与心脏谱系决定的激活 基因，2)CLP-1将前体细胞不可逆转地定位于 3)CLP-1与BMP-2的等级关系， 另一个已知的对承诺的重要发育信号 干细胞移植到心脏谱系。目标3将侧重于描述 控制CLP-1表达所需的启动子元件。申请者将会 检测小鼠CLP-1基因的启动子区域，以确定 在心源性中胚层细胞中定向表达所需的序列(S)。 目标4将试图确定CLP-1在中国的重要性和必要性 活体心脏发育程序的实际阐述。这将是 在基因敲除CLP-1基因后的小鼠中进行评估。 这些小鼠的发育将主要在以下方面进行跟踪 发育中心脏和继发性心脏的形态组织学状况 关于非心脏表型，如果它们出现的话。心脏异常 发展将在分子水平上分析，通过分析 正常心脏遗传标记物的表达可能受 CLP-1转录因子。此外，对转基因植物的生理分析 成年人的心脏将被用来寻找导致 可检测到的心脏功能变化。

DESCRIPTION: (Adapted from the Investigator's Abstract) The applicant hopes to understand the role of cardiogenic transcription factors in the determination and development of cardiac muscle cells. He has identified and cloned a cDNA encoding a cardiac regulatory transcription factor, called Clp-1, that may play an important role in the development of cardiogenic progenitor cells. The stated goal of the current application is to understand the role of Clp-1 in cardiogenesis by addressing when, where and in what capacity Clp-1 performs its function as a regulator of cardiogenesis. The proposal has four specific aims. Aim 1 is directed towards additional characterization of the structural and functional properties of Clp-1 as it relates to its potential cardiogenic regulatory properties. The applicant and his colleagues will attempt to identify the structural domains of Clp-1 that are responsible for binding the B element MEF-2 binding site in the cardiac MLC-2 gene promoter and determine if these same structural domains mediate Clp-1 activation of the cardiac MLC-2 gene in vivo. Aim 2 is directed towards investigating the cellular functions of Clp-1 in order to establish its role as a regulator of cardiac genes and cardiogenic cell differentiation. These studies will focus on determining 1) the potential of Clp-1 to act as a cardiogenic transcriptional activator capable of participating in the activation of cardiac lineage-determining genes, 2) the ability of Clp-1 to irreversibly commit precursor cells to the cardiogenic cell lineage, and 3) the hierarchal relationship of Clp-1 to BMP-2, another developmental signal which is known to be important in commitment of stem cells to the cardiac lineage. Aim 3 will focus on the characterization of promoter elements required for control of Clp-1 expression. The applicant will examine the promoter region of the mouse Clp-1 gene in an effort to define the sequence(s) required for directing expression in cardiogenic mesodermal cells. Aim 4 will attempt to determine the importance and necessity of Clp-1 in the actual elaboration of the cardiogenic developmental program in vivo. This will be evaluated in knockout mice following genetic ablation of the Clp-1 locus. The development of these mice will be followed primarily with respect to the morphological and histological status of the developing heart and secondarily with regard to non-cardiac phenotypes should they arise. Abnormal heart development will be analyzed at the molecular level by assaying for the expression of normal heart genetic markers presumably under the control of the Clp-1 transcription factor. In addition, physiological analysis of transgenic adult hearts will be undertaken to look for subtle perturbations that lead to detectable changes in heart function.