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TUMOR SUPPRESSOR P53 AND THE CNS

肿瘤抑制因子 P53 和中枢神经系统

基本信息

批准号：
2883673
负责人：
STEVEN Scott SCHREIBER
金额：
$ 17.62万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
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：
1997-05-01 至 2001-06-30
项目状态：
已结题

项目摘要

DESCRIPTION: The long-term objective of this proposal is to define mechanisms of abnormal cell cycle regulation leading to selective neuronal degeneration. This will be investigated by in vivo studies of expression, post-translational modifications and biological function of cell cycle regulatory genes in animal models of neuronal apoptosis. Specific Aim 1 will analyze the effects of kainic acid-induced seizures on the post-translational phosphorylation and DNA binding of the p53 transcription factor, a major regulator of the cell cycle that has been implicated in neuronal apoptosis. A solid-phase kinase assay will be used to isolate and measure p53 kinase activity in hippocampal lysates. DNA binding activity will be analyzed by gel mobility shift assay. Specific Aim 2 will analyze the activation of p53-regulated cell cycle genes during adrenalectomy- induced apoptosis of hippocampal granule cells. The cell cycle genes to be studied include cyclin D1, p21 WAF1 and the retinoblastoma susceptibility gene, RB. Wild type mice and mice that are either homozygous (p53-/-) or heterozygous (p53+/-) for the p53 null allele, will be used to elucidate the roles of these genes as effectors of p53-mediated neuronal death. To identify neuronal populations that constitutively express p53, Specific Aim 3 will define regional and cellular patterns of p53 gene expression in the normal postnatal, adult and aged central nervous system. Gene expression will be analyzed by in situ hybridization and immunohistochemistry. The degree of cellular injury will be assessed by histological staining techniques. The results of these studies will add significantly to our knowledge of mechanisms of selective neuronal degeneration related to aberrant cell cycle control, and will provide insight into the pathophysiology of neurodegenerative diseases.

说明：本提案的长期目标是确定细胞周期调节异常导致选择性神经元凋亡的机制退化这将通过体内表达研究来研究，翻译后修饰与细胞周期的生物学功能神经元凋亡动物模型中的调节基因。具体目标1 将分析红藻氨酸诱导的癫痫发作对 p53转录的翻译后磷酸化和DNA结合因子，细胞周期的主要调节因子，神经元凋亡将使用固相激酶测定来分离和测量海马裂解物中的p53激酶活性。 DNA结合活性将通过凝胶迁移率变动分析进行分析。第2章分析肾上腺切除术中p53调节的细胞周期基因的激活- 诱导海马颗粒细胞凋亡。细胞周期基因研究包括细胞周期蛋白D1、p21 WAF 1和视网膜母细胞瘤易感性。基因，RB. 野生型小鼠和纯合子（p53-/-）或 p53无效等位基因的杂合子（p53+/-），将用于阐明这些基因作为p53介导的神经元死亡的效应物的作用。到鉴定组成型表达p53的神经元群体，特异性目的 3将定义p53基因表达的区域和细胞模式，正常的出生后、成年和老年中枢神经系统。基因表达将通过原位杂交和免疫组织化学进行分析。的通过组织学染色评估细胞损伤程度技术. 这些研究的结果将大大增加我们的选择性神经元变性机制的知识，异常细胞周期控制，并将提供深入了解神经退行性疾病的病理生理学。