Using Drosophila to investigate the development and function of nervous systems.

使用果蝇研究神经系统的发育和功能。

• 批准号: RGPIN-2017-05142
• 负责人: vanMeyel, Donald
• 金额: $ 2.91万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2017
• 资助国家: 加拿大
• 起止时间: 2017-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=630087
• 关键词: Using; Drosophila; investigate; development; function

Monoamines are class of neuromodulators that are crucial for a variety of brain functions, including control of mood, movement, cognition, and sleep. Like mammals, the Drosophila nervous system is enriched in monoamines, mainly dopamine, serotonin and octopamine. Distinct populations of monoaminergic neurons release these modulators within anatomically overlapping regions, allowing the brain to integrate environmental cues and regulate animal physiology. Re-uptake of these monoamines then occurs via specific transport proteins to prevent increased extracellular concentration at synapses. During normal cellular metabolism, monoamines are acetylated by the enzyme dopamine N-acetyl transferase (Dat), a Drosophila member of the conserved family of aaNAT (alkylarylamine N-acetyltransferase) proteins. Interestingly, N-acetyl serotonin is a precursor in the synthesis of melatonin, a hormone that induces sleep. We have made a new antibody to Dat and found it to be expressed in some but not all classes of monoaminergic neurons, and likely in neurons expressing other neurotransmitters such as GABA and glutamate. Strikingly, we also found Dat to be expressed in astrocytes, where it is enriched in the fine membranous processes that infiltrate CNS neuropil. The physiological role of astrocytes in monoamine processing is not well understood. We hypothesize that Drosophila astrocytes take up monoamines from synapses and process them via Dat to affect neuromodulation of specific behaviors. In preliminary experiments we find that Dat produced in astrocytes is required for flies to achieve correct levels and timing of sleep. In this grant application, we propose to use genetic approaches to investigate Dat function in Drosophila astrocytes, and to explore with immunohistochemistry, biochemistry and behavioral assays how Dat function might provide selective activity depending on the cell type in which it is expressed. In Aim 1 we will characterize the spatial and temporal patterns of Dat expression in the adult fly brain. In Aim 2 we will measure the biochemical effects of Dat activity in astrocytes and specific neuronal subtypes. In Aim 3 we will determine how Dat influences sleep in Drosophila.

单胺类是一类神经调节剂，对多种大脑功能至关重要，包括控制情绪、运动、认知和睡眠。像哺乳动物一样，果蝇的神经系统富含单胺，主要是多巴胺、血清素和章鱼胺。不同种群的单胺能神经元在解剖学上重叠的区域释放这些调节剂，使大脑能够整合环境信号并调节动物生理。这些单胺的再摄取通过特定的转运蛋白发生，以防止突触细胞外浓度的增加。在正常的细胞代谢过程中，单胺被多巴胺n -乙酰转移酶（Dat）酶乙酰化，多巴胺n -乙酰转移酶（Dat）是果蝇aaNAT（烷基丙胺n -乙酰转移酶）蛋白保守家族的成员。有趣的是，n -乙酰血清素是褪黑素合成的前体，褪黑素是一种诱导睡眠的激素。我们已经制造了一种新的Dat抗体，并发现它在一些但不是所有类型的单胺能神经元中表达，并且可能在表达其他神经递质如GABA和谷氨酸的神经元中表达。引人注目的是，我们还发现Dat在星形胶质细胞中表达，它在浸润中枢神经细胞的细膜突中富集。星形胶质细胞在单胺加工中的生理作用尚不清楚。我们假设果蝇星形胶质细胞从突触中吸收单胺，并通过数据处理它们，从而影响特定行为的神经调节。在初步实验中，我们发现星形胶质细胞中产生的数据是果蝇达到正确睡眠水平和时间所必需的。在这项拨款申请中，我们建议使用遗传学方法来研究果蝇星形胶质细胞中的Dat功能，并通过免疫组织化学、生物化学和行为分析来探索Dat功能如何根据其表达的细胞类型提供选择性活性。在目标1中，我们将描述成年果蝇大脑中数据表达的空间和时间模式。在目标2中，我们将测量Dat活性在星形胶质细胞和特定神经元亚型中的生化效应。在目标3中，我们将确定数据如何影响果蝇的睡眠。