MAP KINASES IN OLIGODENDROCYTE CELL SIGNALING

少突胶质细胞信号传导中的图谱激酶

• 批准号: 6640402
• 负责人: NARAYAN R BHAT
• 金额: $ 24.27万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6640402
• 关键词: biological signal transduction; cell cycle; cell differentiation; cyclin dependent kinase; developmental neurobiology; gel mobility shift assay; gene expression; immunocytochemistry; laboratory rat; mitogen activated protein kinase; myelination; myelinopathy; oligodendroglia; phosphorylation; tissue /cell culture; transfection

DESCRIPTION (provided by applicant): This project is designed to investigate the roles of two related members of the mitogen-activated protein kinase (MAPK) family, i.e., extracellular signal-regulated kinase (ERK) and p38 MAPK in the development and differentiation of oligodendrocytes, the key cells of myelination and demyelination in the CNS. The studies have implications for an understanding of the molecular mechanisms of neural development, myelination and remyelination in demyelinating diseases such as multiple sclerosis. Since the development of oligodendrocytes is stringently controlled by environmental signals, the long-range objective of this research will define signal transduction mechanisms regulating their differentiation and the expression of myelin-specific genes. Primary cultures of rat brain oligodendrocyte progenitors responding to growth/differentiation stimuli will serve as the experimental model to accomplish the following specific aims: * Specific inhibitors of ERK and p38 MAPK will be used to investigate the roles of the two kinases in mitogenic induction and lineage progression (i.e., expression of stage-specific antigens) of oligodendrocyte progenitors as well as in the reciprocal regulation of the levels of key cell cycle regulators, i.e., cyclin D1 (an activator of cyclin-dependent protein kinase, cdk) and p27 (a cdk inhibitor). * The roles of p38 MAPK isoforms and ERKs in oligodendrocyte differentiation will be further characterized using a molecular approach, i.e., transfection with dominant negative and/or constitutively active forms of p38 MAPK isoforms (i.e., alpha and beta) and ERKs (i.e., ERKI and ERK2) as well as their upstream kinases followed by an analysis of the expression of differentiation markers. Putative cytoplastic and nuclear targets (i.e., CREB, ATF2, C/EBP and MEF2C) of p38 MAPK pathway that may signal oligodendrocyte differentiation will be characterized. * p38 MAPK-mediated transcriptional activation of oligodendrocyte-specific genes (i.e., myelin basic protein, protoelipid protein) will be examined using promoter-reporter constructs co-transfected with constitutively active forms of the kinases. Putative MAPK-responsive elements and relevant binding activities will be determined by deletion and gel shift analyses

描述（由申请人提供）：该项目旨在研究有丝分裂原激活蛋白激酶（MAPK）家族的两个相关成员，即外细胞信号调节激酶（ERK）和p38 MAPK在少突胶质细胞的开发和差异化中的p38 MAPK在肌酸和Demyelination of Myelination and Demyelins in of Myelination and Demyelination in of Myelination和Cnss in snss in snss in of p38 MAPK的作用。这些研究对理解神经发育，髓鞘形成和脱髓鞘性疾病（如多发性硬化症）的分子机制有影响。由于少突胶质细胞的发展受到环境信号的严格控制，因此这项研究的远程目标将定义调节其分化的信号转导机制和髓磷脂特异性基因的表达。对生长/分化刺激做出反应的大鼠大脑少生型祖细胞的原发性培养物将作为实现以下特定目标的实验模型： * ERK和p38 MAPK的特定抑制剂将用于研究两个激酶在有丝化诱导和谱系中的作用关键细胞周期调节剂水平的相互调节，即Cyclin D1（Cyclin依赖性蛋白激酶，CDK）和P27（CDK抑制剂）。 * p38 MAPK同工型和Erks在少突胶质细胞分化中的作用将进一步表征，即具有主要的负面和/或组成性活性形式的转染，p38 MAPK同工型（即Alpha）和Erks（即Erks and Erki和Erki和Erki and consears and conseal selys）的作用，以及他们的表现。标记。 p38 MAPK途径的推定细胞塑性和核靶标（即CREB，ATF2，C/EBP和MEF2C）可能会表明少突胶质细胞分化。 * p38 MAPK介导的少突胶质细胞特异性基因（即髓磷脂碱性蛋白，原纤维素蛋白）的转录激活将使用启动子 - 培养剂构建体进行检查。推定的MAPK响应元素和相关的结合活动将通过删除和凝胶转移分析确定