PKA Activation Bypasses the Requirement for UNC-31 in the Docking of Dense Core Vesicles from C. elegans Neurons
PKA Activation Bypasses the Requirement for UNC-31 in the Docking of Dense Core Vesicles from C. elegans Neurons
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DOI：
10.1016/j.neuron.2007.09.015
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发表时间：
2007-11
期刊：
影响因子：
16.2
通讯作者：
Keming Zhou;Yongming Dong;Qian Ge;Dan Zhu;W. Zhou;Xian-guang Lin;T. Liang;Zhengxing Wu;Tao Xu
中科院分区：
文献类型：
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作者：
Keming Zhou;Yongming Dong;Qian Ge;Dan Zhu;W. Zhou;Xian-guang Lin;T. Liang;Zhengxing Wu;Tao Xu
线虫C。ELENGIS为探索突触发生和神经传递的分子基础提供了一个强大的模型系统。然而，缺乏对释放过程的直接功能分析，在很大程度上阻碍了对囊泡胞吐和内吞作用机制的深入了解。优雅女装。我们通过在原代培养中开发包括膜电容和安培测量在内的直接电生理测试来解决这一技术限制。优雅的神经元。此外，我们还成功地利用全内反射荧光显微镜监测了单个致密核心小泡(DCV)的对接和融合。通过这些方法和突变扰动分析，我们提供了直接证据，表明UNC-31是DCV在质膜上对接所必需的。有趣的是，由UNC-31突变引起的DCV对接缺陷可以通过PKA激活完全挽救。我们还证明了UNC-31对于UNC-13介导的DCV胞吐作用的增强是必需的。
The nematodeC. elegansprovides a powerful model system for exploring the molecular basis of synaptogenesis and neurotransmission. However, the lack of direct functional assays of release processes has largely prevented an in depth understanding of the mechanism of vesicular exocytosis and endocytosis inC. elegans. We address this technical limitation by developing direct electrophysiological assays, including membrane capacitance and amperometry measurements, in primary culturedC. elegansneurons. In addition, we have succeeded in monitoring the docking and fusion of single dense core vesicles (DCVs) employing total internal reflection fluorescence microscopy. With these approaches and mutant perturbation analysis, we provide direct evidence that UNC-31 is required for the docking of DCVs at the plasma membrane. Interestingly, the defect in DCV docking caused by UNC-31 mutation can be fully rescued by PKA activation. We also demonstrate that UNC-31 is required for UNC-13-mediated augmentation of DCV exocytosis.