GAD AND GABA-R CANDIDATE GENES FOR EPILEPSY

本提案第一部分中的实验将验证该假设 GABA的结构和调节的变化 谷氨酸脱羧酶(GAD)或GABAA的合成酶 受体多肽是动物惊厥障碍的基础 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 在现有的癫痫小鼠遗传模型中发生了改变， 以及(2)确定GAD或GABA的表达 受体基因在几种遗传和实验中发生了变化 模特或癫痫。 本提案第二部分中提议的实验将尝试 对GAD和GABA受体表达的改变进行编程 细胞系和转基因小鼠中的多肽。第II部 包括三个具体目标：(3)编制高水平的程序 GAD在现有细胞系亚克隆中的表达，(4)至 确定这些工程化细胞中GABA释放的机制 细胞系，以及(5)确定癫痫敏感性是否 GAD或GAD基因表达改变的转基因小鼠的改变 GABA受体。 在某些情况下将GABA激动剂直接注射到大脑中 防止癫痫的传播和细胞死亡。一个长期目标是 在具体目标3和4中提出的实验是 工程化细胞系的生产可以提供持续的 和可调节的GABA来源。这种“设计神经元”可能是 最终在难治性移植治疗中有用 癫痫发作或亨廷顿氏症等神经退行性疾病 疾病。如果我们成功制造出释放GABA的细胞， 无论是根据宪法还是为了应对去极化，我们都将 然后，与其他调查人员合作，解决 实验动物移植中存在的问题及检测 以抑制癫痫发作。

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.