Regulation of PKC alpha in Alcoholic Myocytes

酒精性肌细胞中 PKC α 的调节

• 批准号: 6739095
• 负责人: MICHELE L SOLEM
• 金额: $ 15.7万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6739095
• 关键词: RNase protection assay; alcoholic beverage consumption; alcoholism /alcohol abuse; biological signal transduction; calcium channel; cardiac myocytes; disease /disorder model; enzyme activity; enzyme mechanism; ethanol; gene expression; immunofluorescence technique; immunoprecipitation; insulinlike growth factor; laboratory rat; membrane activity; microarray technology; northern blottings; phosphatidylinositol 3 kinase; phospholipase C; protein biosynthesis; protein kinase C; protein protein interaction; protein tyrosine kinase; western blottings

DESCRIPTION (provided by applicant): This is a revised grant application to study PKC alpha in the alcoholic heart. IGF1 is an essential cytokine that is required for normal development of the heart, and it may play a critical physiological role in the normal adult heart as well as influence various pathological conditions. Our laboratory finds that IGF1 activates PKC alpha in adult rat cardiomyocytes, and this results in PKC alpha-dependent activation of MAP kinase, protein synthesis and gene expression. However, in cardiomyocytes from chronic alcohol-exposed rats, there is reduced activation of PKC alpha by IGF1, loss of IGF1-dependent protein synthesis and a significant increase in the unstimulated (basal) state of the IGF1 receptor. Since chronic alcohol exerts such a profound effect on the IGF1 signaling pathway, an examination of how PKC alpha is involved in these changes may uncover novel cellular mechanisms associated with chronic alcohol-induced damage to the heart. The first aim of this research proposal (1) will be to determine how chronic alcohol alters IGF1-induced PKC alpha activation. What are the changes brought about by chronic alcohol exposure that suppress activation of PKC alpha (e.g. alterations in the IGF1 receptor activation, including the possible involvement of PKC, and the downstream influence on the activities of phospholipase C gamma, PI3 kinase and intracellular tyrosine kinases. The second aim of this proposal (2) will be to study how alterations in IGF1 signaling affect PKC alpha-dependent regulation of protein synthesis and gene expression. Results from cDNA Expression Arrays indicate that the expression of several essential cardiac-specific genes which are involved in excitation-contraction coupling and cell metabolism (e.g. GIRK4, carnitine palmoyltyl transferase, HSP60, G2/M-specific cyclin G), are regulated by PKC alpha-dependent activation by IGF1. This part of the grant proposal will be done in collaboration with the Daniel Baugh Institute at Thomas Jefferson University, using its BioRobotics MicroGridTAS system for microarray analysis of gene expression. The last aim (3) of the proposal will examine how long term alcohol consumption alters the IGF1 signal transduction pathway using the laboratory's model, the chronic alcohol-fed rat fed a diet containing 37% alcohol for various time points of alcohol exposure (e.g. 2 weeks, 2 months, 4 months, 6 months, 10 months). A comparison of the cardiac Ca2+ channel activity, PKC activation and protein synthesis will be made. The objective is to better define the sequence of events that are associated with alcohol-impaired alterations in IGF1 signaling.

描述（由申请人提供）： 这是对酒精心脏中研究PKCα的修订授予申请。 IGF1是心脏正常发育所必需的必不可少的细胞因子，它可能在正常的成人心脏中起关键的生理作用，并且会影响各种病理状况。我们的实验室发现，IGF1在成年大鼠心肌细胞中激活PKCα，这导致PKCα依赖于MAP激酶，蛋白质合成和基因表达的激活。但是，在来自慢性酒精暴露大鼠的心肌细胞中，IGF1的PKCα激活降低，IGF1依赖性蛋白质合成的丧失以及未刺激的IGF1受体状态显着增加（基底）。由于慢性酒精对IGF1信号通路产生了如此深远的影响，因此对PKCα如何参与这些变化的研究可能会发现与慢性酒精引起的心脏损害有关的新型细胞机制。该研究建议（1）的第一个目的是确定慢性酒精如何改变IGF1诱导的PKCα激活。慢性酒精暴露抑制PKCα激活带来的变化是什么（例如，IGF1受体激活的改变，包括PKC的可能参与和下游影响 关于磷脂酶C伽马，PI3激酶和细胞内酪氨酸激酶的活性。该提案的第二个目的（2）将是研究IGF1信号传导的改变如何影响PKCα依赖性蛋白质合成和基因表达的调节。 cDNA表达阵列的结果表明，与激发型偶联和细胞代谢有关的几种必需心脏特异性基因的表达（例如GIRK4，肉碱合理转移酶，HSP60，HSP60，G2/M M特异性cyclin G）受PKC Alpha依赖性激活的调节。赠款提案的这一部分将与托马斯·杰斐逊大学（Thomas Jefferson University）的丹尼尔·鲍尔（Daniel Baugh）研究所合作，使用其生物体微网络系统进行基因表达的微阵列分析。该提案的最后一个目标（3）将检查使用实验室模型的长期饮酒时间改变IGF1信号转导途径，慢性酒精喂养的大鼠为饮食中的饮食喂了37％的酒精饮食（例如2周，2周，2周，2个月，4个月，6个月，6个月，10个月）。将进行心脏Ca2+通道活性，PKC激活和蛋白质合成的比较。目的是更好地定义与IGF1信号中酒精损害变化相关的事件序列。