Stimulus-dependent modifications in astrocyte-derived extracellular vesicle cargo regulate neuronal excitability
Stimulus-dependent modifications in astrocyte-derived extracellular vesicle cargo regulate neuronal excitability
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DOI：
10.1002/glia.23708
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发表时间：
2019-08-30
期刊：
影响因子：
6.2
通讯作者：
Haughey, Norman J.
中科院分区：
文献类型：
--
作者：
Chaudhuri, Amrita Datta;Dasgheyb, Raha M.;Haughey, Norman J.
细胞外囊泡现在已经成为细胞间通讯的关键参与者。这在中枢神经系统中尤其重要，其中胶质神经元串扰有助于维持正常的神经元功能。星形胶质细胞衍生的细胞外囊泡（ADEV）组成性分泌促进神经突起生长和神经元存活。然而，细胞外刺激可以改变ADEV的货物和下游功能。例如，响应于炎症而分泌的ADEV含有货物微RNA和蛋白质，其减少神经突生长、神经元放电并促进神经元凋亡。我们进行了全面的定量蛋白质组学分析，以计数响应于多种刺激而分泌的ADEV的蛋白质组货物。用营养刺激（三磷酸腺苷，ATP）、炎症刺激（IL-1 β）或抗炎刺激（IL-10）刺激大鼠原代星形胶质细胞，并使用连续超离心法收集在2小时时间范围内分泌的细胞外囊泡。使用在不暴露于任何刺激的情况下组成型分泌的ADEV作为对照。使用基于串联质量标签的蛋白质组学平台来鉴定和定量ADEV中的蛋白质。通过免疫途径分析来预测ADEV调控的下游信号事件。我们发现，在响应ATP或IL 10，ADEV包含一组蛋白质，参与增加神经突生长，树突分支，突触传递的调节，并促进神经元存活。相比之下，响应于IL-1 β分泌的ADEV含有调节外周免疫应答和免疫细胞运输至中枢神经系统的蛋白质。
Extracellular vesicles have now emerged as key players in cell-to-cell communication. This is particularly important in the central nervous system, where glia-neuron cross-talk helps maintain normal neuronal function. Astrocyte-derived extracellular vesicles (ADEVs) secreted constitutively promote neurite outgrowth and neuronal survival. However, extracellular stimuli can alter the cargo and downstream functions of ADEVs. For example, ADEVs secreted in response to inflammation contain cargo microRNAs and proteins that reduce neurite outgrowth, neuronal firing, and promote neuronal apoptosis. We performed a comprehensive quantitative proteomic analysis to enumerate the proteomic cargo of ADEVs secreted in response to multiple stimuli. Rat primary astrocytes were stimulated with a trophic stimulus (adenosine triphosphate, ATP), an inflammatory stimulus (IL-1 beta) or an anti-inflammatory stimulus (IL10) and extracellular vesicles secreted within a 2 hr time frame were collected using sequential ultracentrifugation method. ADEVs secreted constitutively without exposure to any stimulus were used a control. A tandem mass tag-based proteomic platform was used to identify and quantify proteins in the ADEVs. Ingenuity pathway analysis was performed to predict the downstream signaling events regulated by ADEVs. We found that in response to ATP or IL10, ADEVs contain a set of proteins that are involved in increasing neurite outgrowth, dendritic branching, regulation of synaptic transmission, and promoting neuronal survival. In contrast, ADEVs secreted in response to IL-1 beta contain proteins that regulate peripheral immune response and immune cell trafficking to the central nervous system.