Science2014-06-10 1:31 PM

3D突触模型揭示突触囊泡再循环中发生了什么 Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins

论文摘要 

Benjamin Wilhelm及其同事从成年大鼠脑中清除了被分离的突触末梢并详细观察了这一膜转运通路,通过该膜转运通路,释放神经递质的突触囊泡被摧毁及重建。研究人员用蛋白印迹和质谱分析来确定有关的蛋白数;用电子显微镜来检测细胞器的数量、大小及位置,及用超高分辨率的荧光显微镜来给蛋白定位。有趣的是,Wilhelm及其同事发现,在突触囊泡再循环的每一步中,每种蛋白的拷贝数是相关的--但它们的数字却在每一步之间有很大的差异。最后,研究人员的这种方法也可能被用于探索其它的细胞转运事件。

Abstract 

Synaptic vesicle recycling has long served as a model for the general mechanisms of cellular trafficking. We used an integrative approach, combining quantitative immunoblotting and mass spectrometry to determine protein numbers; electron microscopy to measure organelle numbers, sizes, and positions; and super-resolution fluorescence microscopy to localize the proteins. Using these data, we generated a three-dimensional model of an “average” synapse, displaying 300,000 proteins in atomic detail. The copy numbers of proteins involved in the same step of synaptic vesicle recycling correlated closely. In contrast, copy numbers varied over more than three orders of magnitude between steps, from about 150 copies for the endosomal fusion proteins to more than 20,000 for the exocytotic ones.

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