Quantitative super-resolution imaging of Bruchpilot distinguishes active zone states.
Quantitative super-resolution imaging of Bruchpilot distinguishes active zone states.
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DOI：
10.1038/ncomms5650
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发表时间：
2014-08-18
影响因子：
16.6
通讯作者：
Kittel, Robert J.
中科院分区：
文献类型：
--
作者：
Ehmann, Nadine;van de Linde, Sebastian;Alon, Amit;Ljaschenko, Dmitrij;Keung, Xi Zhen;Holm, Thorge;Rings, Annika;DiAntonio, Aaron;Hallermann, Stefan;Ashery, Uri;Heckmann, Manfred;Sauer, Markus;Kittel, Robert J.
突触活性区（AZ）的精确分子结构产生了不同的结构和功能AZ状态，从根本上塑造了化学神经传递。然而，阐明在AZ的纳米级蛋白质排列是由传统的光学显微镜衍射有限的分辨率阻碍。在这里，我们介绍了新的方法来量化内源性蛋白质组织在单分子分辨率原位与超分辨率成像直接随机光学重建显微镜（dSTORM）。以果蝇神经肌肉接头（NMJ）为研究对象，发现AZ细胞基质（CAZ）由约137个Bruchpilot（Brp）蛋白组成，其中四分之三的Brp蛋白组成约15个七聚体簇。我们测试CAZ超微结构和神经递质释放特性之间的定量关系，从事果蝇突变体和电生理学。我们的研究结果表明，精确的纳米组织的Brp区分不同的生理AZ状态和链接功能多样化到迄今为止未被识别的神经元梯度的CAZ超微结构。 突触前末梢内的活动区细胞基质（CAZ）中发生复杂的分子相互作用以调节突触可塑性。在这里，作者使用成像技术显示CAZ由平均包含137个Bruchpilot蛋白的单元组成，其中许多蛋白排列成簇。
The precise molecular architecture of synaptic active zones (AZs) gives rise to different structural and functional AZ states that fundamentally shape chemical neurotransmission. However, elucidating the nanoscopic protein arrangement at AZs is impeded by the diffraction-limited resolution of conventional light microscopy. Here we introduce new approaches to quantify endogenous protein organization at single-molecule resolution in situ with super-resolution imaging by direct stochastic optical reconstruction microscopy (dSTORM). Focusing on the Drosophila neuromuscular junction (NMJ), we find that the AZ cytomatrix (CAZ) is composed of units containing ~137 Bruchpilot (Brp) proteins, three quarters of which are organized into about 15 heptameric clusters. We test for a quantitative relationship between CAZ ultrastructure and neurotransmitter release properties by engaging Drosophila mutants and electrophysiology. Our results indicate that the precise nanoscopic organization of Brp distinguishes different physiological AZ states and link functional diversification to a heretofore unrecognized neuronal gradient of the CAZ ultrastructure. Complex molecular interactions occur in the active zone cytomatrix (CAZ) within the presynaptic terminal to regulate synaptic plasticity. Here, the authors use imaging techniques to show that the CAZ is composed of units containing on average 137 Bruchpilot proteins, many of which are arranged into clusters.
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DOI：
10.1126/science.1166673
发表时间：
2009-04-03
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gilestro GF;Tononi G;Cirelli C
通讯作者：
Cirelli C
影响因子：
16.2
作者：
Boyken, Janina;Gronborg, Mads;Chua, John Jia En
通讯作者：
Chua, John Jia En
影响因子：
25
作者：
Guerrero, G;Reiff, DF;Isacoff, EY
通讯作者：
Isacoff, EY
影响因子：
4.8
作者：
Bar-On, Dana;Wolter, Steve;Ashery, Uri
通讯作者：
Ashery, Uri
影响因子：
56.9
作者：
Betzig, Eric;Patterson, George H.;Hess, Harald F.
通讯作者：
Hess, Harald F.