GAD AND GABA-R CANDIDATE GENES FOR EPILEPSY

本提案第一部分的实验将检验这一假设 GABA的结构和调节的改变 合成酶，谷氨酸脱羧酶（GAD），或GABAA 受体多肽是动物癫痫病的基础 癫痫模型 第一部分包括两个具体目标：（1）确定是否 编码GAD或GABA受体多肽的基因之一是 在一个现有的癫痫遗传小鼠模型中发生了改变， （2）检测GAD或GABA的表达 受体基因在几种遗传和实验中发生改变， 模型或癫痫。 本提案第二部分提出的实验将试图 编程GAD和GABA受体的表达改变， 在细胞系中和在转基因小鼠中表达多肽。 第二部分 具体包括三个目标：（3）规划高水平的 GAD在现有细胞系的亚克隆中的表达，（4） 确定GABA释放的机制，在这些工程 细胞系，和（5）以确定癫痫易感性是否是 在GAD表达改变的转基因小鼠中发生改变，或 GABA受体。 在某些情况下，将GABA激动剂直接注射到大脑中 防止癫痫蔓延和细胞死亡。 长期目标是 具体目标3和4中提出的实验是 生产工程细胞系，可以提供持续的 和可调节的GABA来源。 这种“设计神经元”可能是 最终可用于移植治疗顽固性 癫痫发作或神经退行性疾病如亨廷顿病 疾病 如果我们成功制造出释放GABA的细胞， 无论是组成性的还是对去极化的反应，我们将 然后，与其他研究人员合作，解决 移植到实验动物和测试中的问题 用于抑制癫痫发作。

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.