Molecular neurogenetics of seizure mutants in Drosophila

描述（由申请人提供）：果蝇行为突变的表征已经确定了过多的突触蛋白和离子通道。此外，最近的实验和基因组比较已经证明了相同的蛋白质在许多人类神经系统疾病中的作用。我建议在果蝇中使用正向和反向遗传方法来阐明神经系统疾病（如癫痫和涉及异常神经元高兴奋性的相关疾病）的细胞和分子机制。Aim 1提出了一种反向遗传学方法来研究KCNQ K+通道在体内的作用，KCNQ K+通道是神经元兴奋性的调节剂，与癫痫、心律失常和耳聋直接相关。KCNQ通道的时空表达模式将被确定。KCNQ通道的结构-功能分析将通过生成各种位点定向突变动物来进行。这些KCNQ突变果蝇将在行为和生理上进行表征，以确定该通道在神经元信号传导、可塑性和神经疾病中的作用。目的2提出了一种正向遗传学方法来表征三种具有行为和生理癫痫发作的新型过度兴奋突变体，随后是瘫痪。这些受影响的基因已被很好地定位，并将被识别和克隆。此外，肌肉和神经元功能的详细生理特征将被完成，以确定这些突变体中行为检测的具体机制。

DESCRIPTION (provided by applicant): Characterization of behavioral mutants in Drosophila has identified a plethora of synaptic proteins and ion channels. Furthermore, recent experiments and genomic comparisons have demonstrated the role of the same proteins in numerous human neurological diseases. I propose to use forward and reverse genetic approaches in Drosophila to elucidate the cellular and molecular mechanisms underlying neurological diseases such as epilepsy and related disorders that involve aberrant neuronal hyperexcitability. Aim 1 proposes a reverse genetics approach to study the in vivo role of the KCNQ K+ channel, a regulator of neuronal excitability, which has been directly linked to epilepsy, heart arrhythmia and deafness. The temporal and spatial expression pattern of the KCNQ channel will be determined. A structure-function analysis of the KCNQ channel will be performed by generating various site-directed mutant animals. These KCNQ mutant flies will be characterized behaviorally and physiologically to determine the role of this channel in neuronal signaling, plasticity and neurological disorders. Aim 2 proposes a forward genetics approach to characterize three novel hyperexcitable mutants with behavioral and physiological seizures followed by paralysis. These affected genes are well mapped and will be identified and cloned. In addition, detailed physiological characterization of muscle and neuronal function will be done to determine the specific mechanisms underlying the behavioral detects in these mutants.