Systematic discovery of Rab GTPases with synaptic functions in Drosophila.
Systematic discovery of Rab GTPases with synaptic functions in Drosophila.
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DOI：
10.1016/j.cub.2011.08.058
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发表时间：
2011-10-25
期刊：
影响因子：
9.2
通讯作者：
Hiesinger, P. Robin
中科院分区：
文献类型：
--
作者：
Chan, Chih-Chiang;Scoggin, Shane;Wang, Dong;Cherry, Smita;Dembo, Todd;Greenberg, Ben;Jin, Eugene Jennifer;Kuey, Cansu;Lopez, Antonio;Mehta, Sunil Q.;Perkins, Theodore J.;Brankatschk, Marko;Rothenfluh, Adrian;Buszczak, Michael;Hiesinger, P. Robin
神经元需要高度专门化的细胞膜运输，尤其是突触。RAB GTP酶被认为是所有细胞膜转运的主要调节者，只有极少数的RAB具有已知的神经元特异性功能。在这里，我们首次系统地描述了Rab GTP酶在一个有大脑的生物体中的神经元表达、亚细胞定位和功能。我们报告了一个令人惊讶的发现，所有果蝇兔子中有一半专门或主要在大脑中不同的神经元亚群中发挥作用。此外，对GTP/GDP结合状态的功能分析表明，这些神经元RAB几乎只活跃于突触，并且大多数突触RAB专门标记突触循环的内体隔室。我们的图谱分析策略是基于大基因组片段中的Gal4敲入，这些大基因组片段被额外设计成通过末端同源重组产生突变。我们构建了36个大的基因组靶向载体和25个Rab基因的转基因Rab-Gal4果蝇品系。突触Rab27的原理证明敲除揭示了与其细胞特异性表达相匹配的睡眠表型。我们的发现表明，多达一半的果蝇兔发挥着特殊的突触功能。本文提供的工具可以对这些RAB进行系统的功能研究，并提供了一种适用于果蝇任何大基因家族的方法。
Neurons require highly specialized intracellular membrane trafficking, especially at synapses. Rab GTPases are considered master regulators of membrane trafficking in all cells and only very few Rabs have known neuron-specific functions. Here, we present the first systematic characterization of neuronal expression, subcellular localization and function of Rab GTPases in an organism with a brain. We report the surprising discovery that half of all Drosophila Rabs function specifically or predominantly in distinct subsets of neurons in the brain. Furthermore, functional profiling of the GTP/GDP-bound states reveals that these neuronal Rabs are almost exclusively active at synapses and the majority of these synaptic Rabs specifically mark synaptic recycling endosomal compartments. Our profiling strategy is based on Gal4 knock-ins in large genomic fragments that are additionally designed to generated mutants by ends-out homologous recombination. We generated 36 large genomic targeting vectors and transgenic rab-Gal4 fly strains for 25 rab genes. Proof-of-principle knock-out of the synaptic rab27 reveals a sleep phenotype that matches its cell-specific expression. Our findings suggest that up to half of all Drosophila Rabs exert specialized synaptic functions. The tools presented here allow systematic functional studies of these Rabs and provide a method that is applicable to any large gene family in Drosophila.
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影响因子：
64.5
作者：
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通讯作者：
Freeman, M
影响因子：
56.9
作者：
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DOI：
10.1073/pnas.0535280100
发表时间：
2003-03-04
影响因子：
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作者：
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通讯作者：
Golic, KG
DOI：
10.1073/pnas.84.23.8210
发表时间：
1987-12-01
影响因子：
11.1
作者：
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通讯作者：
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影响因子：
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作者：
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