GAD AND GABA-R CANDIDATE GENES FOR EPILEPSY

GAD 和 GABA-R 癫痫候选基因

• 批准号: 3404467
• 负责人: ALLAN J TOBIN
• 金额: $ 23.19万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-04-01 至 1993-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3404467
• 关键词: GABA receptor; biological models; epilepsy; gamma aminobutyrate; genetic mapping; genetic models; genetically modified animals; glutamate decarboxylase; laboratory mouse; molecular cloning; receptor expression; tissue /cell culture

The experiments in Part I of this proposal will test the hypothesis that alterations in the structure and regulation of the GABA synthetic enzyme, glutamate decarboxylase (GAD), or of GABAA receptor polypeptides underlie the seizure disorders in animal models of epilepsy. Part I encompasses two specific aims: (1) to determine whether one of the genes encoding GAD or GABA receptor polypeptides is altered in one of the existing genetic mouse models of epilepsy, and (2) to determine whether the expression of GAD or GABA receptor genes is altered in several genetic and experimental models or epilepsy. The experiments proposed in Part II of this proposal will attempt to program the altered expression of GAD and GABA receptor polypeptides in cell lines and in transgenic mice. Part II encompasses three specific aims: (3) to program high level expression of GAD in subclones of existing cell lines, (4) to determine the mechanism of GABA release in these engineered cell lines, and (5) to determine whether seizure susceptibility is altered in transgenic mice with altered expression of GAD or GABA receptors. Direct injection of GABA agonists into the brain in some cases prevents seizure propagation and cell death. A long term goal of the experiments proposed in specific aims #3 and 4 is the production of engineered cell lines that could provide a continuing and regulatable source of GABA. Such "designer neurons" may be ultimately useful in transplantation therapies for intractable seizures or for such neurodegenerative diseases as Huntington's disease. If we succeed in producing cells that release GABA, either constitutively or in response to depolarization, we will then, in collaboration with other investigators, address the problems of transplantation into experimental animals and testing for seizure suppression.

本提案第一部分的实验将检验假设 GABA 结构和调节的改变 合成酶、谷氨酸脱羧酶 (GAD) 或 GABAA 受体多肽是动物癫痫发作的基础 癫痫模型。 第一部分包含两个具体目标：(1) 确定是否 编码GAD或GABA受体多肽的基因之一是 在一种现有的癫痫遗传小鼠模型中进行了改变， (2)判断是否表达GAD或GABA 受体基因在一些遗传和实验中被改变 模型或癫痫。 本提案第二部分中提出的实验将尝试 编程改变 GAD 和 GABA 受体的表达 细胞系和转基因小鼠中的多肽。 第二部分 包括三个具体目标：（3）规划高水平 GAD 在现有细胞系亚克隆中的表达，(4) 确定这些工程中 GABA 释放的机制 细胞系，以及（5）确定癫痫易感性是否 GAD 表达改变的转基因小鼠发生改变或 GABA 受体。 在某些情况下将 GABA 激动剂直接注射到大脑中 防止癫痫发作传播和细胞死亡。 长期目标是 具体目标#3和4中提出的实验是 生产可以提供持续的工程细胞系 和可调节的 GABA 来源。 这种“设计神经元”可能是 最终可用于顽固性移植治疗 癫痫发作或亨廷顿舞蹈症等神经退行性疾病 疾病。 如果我们成功地生产出释放 GABA 的细胞 无论是本质上还是为了应对去极化，我们将 然后，与其他研究人员合作，解决 实验动物移植及测试问题 用于抑制癫痫发作。