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NEGATIVE CELL CYCLE REGULATION IN CARDIAC MUSCLE CELLS

心肌细胞的负细胞周期调节

基本信息

批准号：
2029901
负责人：
HONG ZHU
金额：
$ 3.62万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-01-20 至 1997-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2029901
关键词：
age difference antisense nucleic acid cell cycle cell differentiation cell growth regulation cell proliferation confocal scanning microscopy developmental genetics gene expression heart ventricle immunofluorescence technique intracellular transport laboratory rat myocardium myogenesis northern blottings transfection yeast two hybrid system

项目摘要

DESCRIPTION (adapted from the applicant's abstract): The long-term goal of this project is to understand the molecular mechanisms that regulate the permanent withdrawal of ventricular cardiomyocytes from the cell cycle during terminal differentiation. The applicant has isolated a gene from juvenile rat ventricles by cDNA subtraction with fetal ventricular cDNA. Northern blot analysis shows expression of the gene in neonatal, juvenile, and adult ventricles but not in fetal ventricles. In neonates, its expression is higher in ventricles than other organs. The deduced amino acid sequence shares about 50% similarity to Growth Arrest-Specific gene 1 (gas1) which suppresses mitosis. Hence, the isolated gene is named c-gas1 for cardiac gas1. Transient expression of c-gas1 in fetal ventricular myocytes results in a significant reduction in DNA synthesis. The applicant hypothesizes that c-gas1 is a developmentally regulated gene which suppresses ventricular myocyte proliferation during and after terminal differentiation. To test this hypothesis, the following specific aims are proposed: 1) To determine the cellular localization of c-gas1 protein in ventricular myocytes by immunofluorescence staining and confocal microscopy. 2) To determine whether abolishing the expression of c-gas1 by antisense oligonucleotides will permit terminally differentiated ventricular myocytes to enter the cell cycle. 3) To identify the cellular factors in terminally differentiated ventricular myocytes that directly interact with c-gas1 protein by the yeast double-hybrid system. 4) to examine the effect of overexpressing c-gas1 in fetal ventricular myocytes and the effect of inactivation of both alleles of the c-gas1 gene on myocardial growth and development.

描述(改编自申请人的摘要)：长期目标这个项目是为了了解调节细胞周期的分子机制。心肌细胞从细胞周期中永久退出在顶端分化过程中。申请人已经分离出一个基因幼年大鼠脑室与胎儿脑室cDNA差减。 Northern印迹分析表明该基因在新生儿、幼年、和成人脑室，但不在胎儿脑室。在新生儿中，它的脑室的表达高于其他器官。推导出的氨基酸酸序列与生长停滞特异基因1有约50%的相似性 (Gas1)抑制有丝分裂。因此，分离到的基因被命名为c-Gas1。心脏Gas1。C-Gas1在胎儿脑室的瞬时表达肌细胞导致DNA合成显著减少。申请人假设c-Gas1是一个发育受调控的基因抑制终末期和终末期后的心肌细胞增殖差异化。为了验证这一假设，以下是具体目标建议：1)确定c-Gas1蛋白在细胞内的定位。用免疫荧光染色和共聚焦显微镜观察心肌细胞。 2)确定是否反义取消c-Gas1的表达寡核苷酸将允许终末分化的心室肌细胞进入细胞周期。3)确定终末期的细胞因子与c-Gas1直接相互作用的分化的心室肌细胞蛋白质的酵母双杂交系统。4)检查以下项目的效果 C-Gas1在胎儿心肌细胞中的过表达及其对心肌细胞生长的影响 C-Gas1基因两种等位基因失活对心肌生长和心肌功能的影响发展。