TRANSGENIC CATACHOLAMINERGIC CELL LINES

转基因儿茶酚胺能细胞系

• 批准号: 3416542
• 负责人: Dona M Chikaraishi
• 金额: $ 22.15万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-20 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3416542
• 关键词: GABA receptor; catecholamines; cell line; chromaffin cells; corpus striatum; developmental neurobiology; diphtheria toxin; dopamine receptor; gene expression; genetic enhancer element; genetic promoter element; genetic regulation; genetic regulatory element; genetically modified animals; glutamate receptor; inbreeding; laboratory mouse; neurons; neuropeptide receptor; oncogenes; reporter genes; substance P; thymidine kinase; tissue /cell culture; transfection; tyrosine 3 monooxygenase; xenotransplantation

Very few neuronal cell lines exhibit differentiated properties of mature neurons. Most existing lines are immature and neuroblast-like, which limits their usefulness. Often the identity of the parental cell type from which the lines arose is unknown. We propose to derive cultured cell lines of catecholaminergic neurons or adrenal medullary cells from tissues of transgenic (TG) mice in which oncogenes are expressed from the rat tyrosine hydroxylase (TH) promoter. Initially we will determine which regulatory elements are necessary for cell specific expression of tyrosine hydroxylase in transgenic mice using both oncogene and non-oncogene reporter genes. We are particularly interested in determining if different TH expressing cells (CNS vs PNS or dopaminergic vs noradrenergic CNS neurons) require different enhancer elements. Transgenic animals will be generated with the appropriate TH enhancer-oncogene constructs and catecholaminergic tissues from oncogene-expressing animals will be cultured. Cell lines will be selected and characterized for a variety of neuronal specific traits. If necessary animals bearing several different oncogenes will be breed to increase the efficiency of immortalization. Lastly, TH+ cells will be ablated during development by targeting the expression of the diphtheria toxin gene or the Herpes simplex virus thymidine kinase gene to catecholaminergic cells in transgenic mice.

很少有神经元细胞系具有分化特性 成熟的神经元。 大多数现有的线不成熟和神经细胞状， 这限制了他们的实用性。 通常是父母细胞的身份 线产生的类型是未知的。 我们建议得出 儿茶酚胺能神经元或肾上腺髓质的培养细胞系 来自癌基因的转基因（TG）小鼠组织的细胞 用大鼠酪氨酸羟化酶（Th）启动子表达。 最初，我们将确定哪些监管要素是 酪氨酸羟化酶在细胞特异性表达中所必需的 使用癌基因和非癌基因报告基因的转基因小鼠。 我们 特别有兴趣确定是否表达不同 细胞（CNS与PNS或多巴胺能与去甲肾上腺素能CNS神经元）需要 不同的增强子元素。 转基因动物将与适当的TH产生 从 将培养表达致癌基因的动物。 细胞系将是 选择并针对各种神经元特异性特征进行特征。 如果 带有几种不同发病基因的必要动物将被繁殖到 提高永生化的效率。 最后，在开发过程中将消融Th+细胞 靶向白喉毒素基因或疱疹的表达 单纯形病毒胸苷激酶基因至儿茶酚胺能细胞中的细胞 转基因小鼠。