RPG: Regulation of Neurotransmitter Transporters in Drosophila

9407849 Neckameyer摘要突触的传递严重依赖于突触裂隙处神经递质的浓度。当神经递质在裂隙处释放时，神经递质转运体有助于将递质移入突触前终末。这限制了神经递质活动的持续时间，并被认为在维持各种突触的神经递质可用性方面发挥着重要作用。调节神经递质转运体活性的确切分子机制才刚刚开始阐明。此外，这些工作还没有解决在体内干扰这些机制的发育和功能意义。这项应用建议建立在我们最近克隆的Drosphila神经递质转运体cDNA的基础上，以检测这些重要蛋白质的调节。选择果蝇使我们能够在这些重要问题上发挥这个模型系统的一些独特优势(例如，在产生转基因动物和特定于遗传位点的突变方面的便利)。这些研究将利用重组DNA方法来(1)确定到目前为止分离的每个转运蛋白的特定神经递质底物，以及(2)启动转运蛋白基因座的分子遗传学分析。这些初步研究将为未来的实验奠定基础，这些实验将通过将改变的转运蛋白基因引入Drosphila的基因组来评估干扰重摄取调节机制在体内的发育和功能后果。这些研究将增加我们对神经递质转运体在突触传递中所起作用的理解。

9407849 Neckameyer ABSTRACT Synaptic transmission is critically dependent on the concentrations of neurotransmitter at the synaptic cleft. Upon release of the neurotransmitter at the cleft, neurotransmitter transporters contribute to the removal of transmitter into the presynaptic terminal. This limits the duration of the neurotransmitter action, and is thought to play a major role in maintaining neurotransmitter availability at a variety of synapses. The precise molecular mechanisms which regulate the activity of the neurotransmitter transporters are only beginning to be elucidated. Moreover, such work has not yet addressed the developmental and functional significance of perturbing these mechanisms in vivo. This application proposes to build upon our recent cloning of Drosphila neurotransmitter transporter cDNAs to examine the regulation of these important proteins. The choice of Drosophila enables us to bring to bear some of the unique advantages of this model system (such as the facility in generating both transgenic animals as well as mutations specific to genetic loci) on these important questions. These studies will take advantage of recombinant DNA methodology to (1) identify the specific neurotransmitter substrate for each of the transporters isolated thus far, and (2) to initiate a molecular genetic analysis of the transporter loci. These initial studies will lay the groundwork for future experiments that will assess the developmental and functional consequences in vivo of perturbing reuptake regulatory mechanisms by introducing altered transporter genes into the genome of Drosphila. These studies will increase our comprehension of the role that neurotransmitter transporters play in synaptic transmission.