M4 MUSCARINIC RECEPTOR MRNA STABILITY

描述：毒扁豆碱受体是许多 包括认知和记忆在内的生理过程。初步数据 表明编码不同M受体亚型的基因可以 实行差别化监管。在克隆性PC12细胞系中，M4受体 NGF处理可以稳定和上调mRNAs。更好的 对这一现象的理解可能与理解基础有关 阿尔茨海默病中M4受体上调的研究。既有药理学上的 在这些研究中将采用分子方法。 具体目标是：1.确定信号转导途径 调节M4受体mRNA的稳定性。对M4受体基因表达的影响 关键蛋白的药理和遗传操纵的稳定性 NGF受体的信号转导途径有待进一步研究。 2.鉴定3‘-非翻译区及其侧翼核苷酸 调节M4受体mRNA稳定性变化的序列。删除 M4受体mRNA的嵌合cDNA构建将被产生并 转染PC12克隆细胞定位顺式作用元件 控制转录稳定性。3.确定反式作用因素的特征 参与调节M4受体mRNA的稳定性。这一部分将重点介绍 M4受体mRNA与M4受体相互作用的检测与表征 调节反式作用的蛋白质因子。

DESCRIPTION: Muscarinic receptors are key participants in many physiological processes including cognition and memory. Preliminary data indicate that genes encoding the different muscarinic receptor subtypes can be differentially regulated. In the clonal PC12 cell line, m4 receptor mRNAs are stabilized and up-regulated by treatment with NGF. A better understanding of this phenomenon may be relevant to understanding the basis of m4 receptor upregulation in Alzheimer's disease. Both pharmacological and molecular approaches will be employed in these studies. The specific aims are: 1. To identify the signal transduction pathways modulating m4 receptor mRNA stability. The effects on m4 receptor mRNA stability by pharmacological and genetic manipulation of key proteins in the signal transduction pathway of NGF receptor will be studies. 2. To characterize the 3'-untranslated region and flanking nucleotides sequences regulating changes in m4 receptor mRNA stability. Deletion of chimeric cDNA constructs of the m4 receptor mRNA will be generated and transfected into PC12 clonal cell line to localize cis-acting elements controlling transcript stability. 3. To characterize trans-acting factors involved in modifying m4 receptor mRNA stability. This section will focus on detecting and characterizing interactions between m4 receptor mRNA and the regulatory trans-acting protein factors.