Real-time measurements of neurotransmission in Drosophilia melanogaster

 描述（由申请人提供）：模型系统用于了解神经传递是如何调节的，以及它在疾病期间是如何发生故障的。果蝇是一种很有吸引力的模式生物，因为它很容易改变基因，繁殖速度快，价格低廉，而且行为复杂。果蝇有许多类似于哺乳动物的神经递质系统，包括多巴胺能和多巴胺能系统。虽然多巴胺和5-羟色胺的测量已在幼虫果蝇腹神经索，没有测量内源性释放已在成年苍蝇的大脑。成年果蝇有一个发育完全的神经系统，将是一个很好的模型系统，用于研究神经递质在精神疾病中的调节。我实验室的长期目标是开发新的分析工具，以了解生物系统中的实时神经传递。本项目的目标是描述成年和幼虫果蝇特定神经元中多巴胺和5-羟色胺信号的调节。具体来说，我们将确定哪些血清素受体作为自动受体，以及多巴胺是如何调节环指蛋白11（RNF 11），NF-κ B B通路的调节剂。核心假设是神经递质调节随年龄和神经元的变化而变化。目的1是比较多巴胺释放在离散地区的幼虫和成人中枢神经系统。将通过光学激活基因插入的CsChrimson来诱发释放，并将在成人中研究衰老对多巴胺释放的影响。在目标2中，我们将使用RNF 11的RNAi敲低来探索RNF 11对多巴胺的调节。将在RNF 11 RNAi果蝇中研究D2自身受体拮抗剂抗精神病药的作用，以研究RNF 11影响多巴胺自身受体调节的程度。这将是RNF 11对任何物种多巴胺信号传导影响的首次研究。在目标3中，我们将研究在幼虫（前脑和VNC）和成虫（蘑菇体，VNC）中刺激的5-羟色胺释放和再摄取。血清素系统是许多精神疾病治疗的目标，因此了解苍蝇中的血清素系统将使其成为精神疾病的模型生物体。最后，对于目标4，我们将确定血清素自身受体对血清素的影响。 利用药理学和遗传学方法在幼虫和成虫中进行调节。这项工作的预期成果是更好的方法，在特定的果蝇neurophilis和了解整个果蝇发育过程中多巴胺和5-羟色胺释放的调节神经传递的特征。这项研究意义重大，因为它将提供必要的神经递质信号传导的基础知识，以使用果蝇可用的强大遗传工具来研究特定基因在控制神经传递中的作用。未来的研究可以研究多巴胺能和多巴胺能神经传递如何控制成年果蝇的行为或疾病功能。

 DESCRIPTION (provided by applicant): Model systems are used to understand how neurotransmission is regulated and how it malfunctions during diseases. Drosophila, the fruit fly, is an attractive model organism because it is easy to genetically alter, fast to breed, inexpensive, and has complex behaviors. Drosophila has many neurotransmitter systems similar to mammals, including dopaminergic and serotonergic systems. While dopamine and serotonin measurements have been made in the larval Drosophila ventral nerve cord, no measurements of endogenous release have been made in the adult fly brain. Adult flies have a fully developed nervous system and would be a good model system for studying the regulation of neurotransmitters in mental health diseases. The long-term goal of my lab is to develop new analytical tools to understand real-time neurotransmission in biological systems. The goal of this project is to characterize the regulation of dopamine and serotonin signaling in specific neuropil of the adult and larval Drosophila. Specifically, we will determine which serotonin receptors act as auto receptors and how dopamine is regulated by RING finger protein 11 (RNF11), a modulator of the NF-B pathway. The central hypothesis is that neurotransmitter regulation varies with age and by neuropil. Aim 1 is to compare dopamine release in discrete areas of the larval and adult CNS. Release will be evoked by optically activating genetically-inserted CsChrimson and the effect of aging on dopamine release will be studied in the adult. In Aim 2, we will explore the regulation of dopamine by RNF11 using RNAi knockdown of RNF11. The effect of D2 auto receptor antagonist antipsychotics will be studied in RNF11 RNAi flies, to investigate the extent to which RNF11 affects auto receptor regulation of dopamine. These will be the first studies of the effect of RNF11 on dopamine signaling in any species. In Aim 3, we will investigate stimulated serotonin release and reuptake in larvae (protocerebrum and VNC) and adults (mushroom bodies, VNC). The serotonin system is a target of many treatments for mental illness, so understanding it in flies will enable its use as a model organism of mental diseases. Finally, for Aim 4, we will determine the effect of serotonin auto receptors on serotonin regulation in larvae and adults using both pharmacological and genetic approaches. The expected outcomes of this work are better methods for characterizing neurotransmission in specific Drosophila neuropil and an understanding of the regulation of dopamine and serotonin release throughout Drosophila development. The research is significant because it will provide foundational knowledge of neurotransmitter signaling necessary to use the powerful genetic tools available for Drosophila to study the effects of specific genes in controlling neurotransmission. Future studies could examine how dopaminergic and serotonergic neurotransmission control behaviors or function in diseases in adult Drosophila.