Lymphoid Transcription Factor in Striatal Development

纹状体发育中的淋巴转录因子

• 批准号: 6536167
• 负责人: Denes V. Agoston
• 金额: $ 7.41万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6536167
• 关键词: cell differentiation; corpus striatum; dopamine receptor; enkephalins; gel mobility shift assay; gene expression; genetic mapping; genetic markers; immunocytochemistry; in situ hybridization; laboratory rat; neurons; phenotype; polymerase chain reaction; transcription factor; western blottings

DESCRIPTION: (Provided by Applicant) Striatal neurons are involved in controlling movement and also in mediating reward, motivation and social behavior. At the level of the nucleus the differentiation of progenitors into different striatal neuronal phenotypes that mediate striatal functions is regulated by transcription factors. Currently, the coordinated and cooperative interactions of transcription factors required for normal striatal development is not well understood. A combination of Western-analysis, RT-PCR, immunohistochemistry and electrophoretic mobility shift assays (EMSA) demonstrated that the lymphoid zinc-finger transcription factor Ikaros, which controls T- and B-cell differentiation is specifically expressed in the developing striatum. A transient transfection assay in differentiating striatal neurons indicated that Ikaros activity is required for the developmental expression of the enkephalin (ENK) gene that encodes one of the most abundant striatal peptide neurotransmitters. The hypothesis of this application is that Ikaros regulates striatal differentiation by regulating the entry of progenitors into the differentiation pathway. The objective of this application is to map Ikaros expression in the developing striatum and to analyze the effect of its altered availability on striatal differentiation. The spatiotemporal pattern of Ikaros expression will be determined at various stages of striatal differentiation in the rat. Ikaros positive cells will be analyzed for proliferation (BrdU incorporation) and for the expression of selected developmental and phenotypic markers, including nestin, MAP2, NF68, ENK, substanceP/Neurokinin, dynorphin, D1 and D2 dopamine receptors, calbindin D-28 and DARRP-32 by immunohistochemistry and by in situ hybridization histochemistry. This information will provide information about the spatiotemporal distribution, developmental status and phenotype of Ikaros expressing cells. The availability of Ikaros will be altered by antisense oligonucleotide treatment or by overexpression in embryonic striatal cultures at various stages of differentiation. Following treatments changes in the expression of markers listed above and the number of apoptotic (TUNEL+) cells will be determined by immunohistochemistry, in situ hybridization histochemistry and TUNEL assay. These experiments will provide information about the role of Ikaros in striatal development. The proposed experiments will help to design future in vivo studies using animal models and in vitro studies focussing on the role of Ikaros in chromatin remodeling implicated in the development lymphoid system. Our studies will advance our knowledge about the molecular requirements for normal striatal development and contribute to a better understanding of developmental brain disorders.

描述：（由申请人提供）纹状体神经元参与 控制运动并调节奖励、动机和社交 行为。 在细胞核水平上，祖细胞分化为 介导纹状体功能的不同纹状体神经元表型是 受转录因子调节。 目前，经协调和 正常纹状体所需的转录因子的协作相互作用 发展还不是很了解。 结合西方分析， RT-PCR、免疫组织化学和电泳迁移率变动分析 (EMSA) 证明淋巴锌指转录因子 Ikaros 控制 T 细胞和 B 细胞分化的特异性表达于 发育中的纹状体。 瞬时转染实验在分化中的应用 纹状体神经元表明 Ikaros 活性是 编码其中一种的脑啡肽 (ENK) 基因的发育表达 最丰富的纹状体肽神经递质。 对此的假设 应用是Ikaros通过调节纹状体分化来调节 祖细胞进入分化途径。 的目标 该应用程序旨在绘制发育中纹状体中的 Ikaros 表达图并 分析其可用性改变对纹状体分化的影响。 Ikaros 表达的时空模式将在不同的时间点确定 大鼠纹状体分化的阶段。 Ikaros 阳性细胞将 分析增殖（BrdU 掺入）和表达 选定的发育和表型标记，包括巢蛋白、MAP2、NF68、 ENK、P 物质/神经激肽、强啡肽、D1 和 D2 多巴胺受体、钙结合蛋白 通过免疫组织化学和原位杂交检测 D-28 和 DARRP-32 组织化学。 此信息将提供有关 Ikaros的时空分布、发育状况和表型 表达细胞。 Ikaros 的可用性将因反义而改变 寡核苷酸治疗或通过在胚胎纹状体培养物中过度表达 处于分化的各个阶段。 以下治疗方法的改变 上述标记物的表达和凋亡 (TUNEL+) 细胞的数量 通过免疫组化、原位杂交等方法确定 组织化学和TUNEL 测定。 这些实验将提供信息 关于 Ikaros 在纹状体发育中的作用。 拟议的实验 将有助于设计未来使用动物模型和体外的体内研究 研究重点是 Ikaros 在染色质重塑中的作用 淋巴系统的发育。 我们的研究将增进我们的知识 正常纹状体发育的分子需求并有助于 更好地了解发育性大脑疾病。