A connectivity-based synaptome

描述（由申请人提供）：突触病理学常见于各种神经退行性和神经发育疾病。对于许多这样的疾病，包括精神分裂症和自闭症谱系障碍，突触组成的变化被怀疑与一些最具破坏性的症状有因果关系。通过电子显微镜观察，大脑中的突触共享关键的结构蛋白，这一点可以通过它们相似的、容易识别的结构来证明，但它们也是非常异质的：已知只有少数突触成分是所有突触共享的。突触前膜与突触后膜之间的相互作用非常强，难以分离。这种相互作用可以承受组织分离，并且在过去几十年中已经被用于纯化突触体：突触前至突触后粘连和重新密封以形成封闭的突触细胞器的附着膜。目前的蛋白质组学研究已利用突触体或亚组分的准备，以评估突触的组成。虽然这支持了使用突触体进行基于蛋白质组学的比较的可行性，但由此产生的大型数据集并不是很有用，因为它们反映了包含巨大异质突触群体的高度复杂的起始材料。突触异质性主要基于连接性的差异，但目前还没有可用的方法可用于询问和分析识别的突触群体之间的差异。在这里，我们提出了一种新的和简单的方法的基础上，哺乳动物的GFP重建突触合作伙伴的技术，这将使健康和疾病的突触群体之间的蛋白质组比较。我们已经组建了一个团队来测试这一点使用定义明确的突触回路。这种方法将是一个重大的进步，因为它将允许快速，大规模的评估特定的突触群体，可以从复杂的电路。

DESCRIPTION (provided by applicant): Synapse pathology is common to a variety of neurodegenerative and neurodevelopmental diseases. For many such disorders, including schizophrenia and autism spectrum disorders, changes in synapse composition are suspected to be causally related to some of the most devastating symptoms. Synapses throughout the brain share key structural proteins evidenced by their similar, readily identified structure as see through an electron microscope, but they are also very heterogeneous: only a handful of synapse components is known to be shared by all synapses. Presynaptic membranes adhere to postsynaptic membranes in an exceptionally strong interaction, resistant to disassembly. This interaction can withstand tissue fractionation and has been exploited over the past several decades to purify synaptosomes: pre- to postsynaptic adhesions and attached membranes that re-seal to make an enclosed synaptic organelle. Current proteomics studies have utilized synaptosomes or subfractions of such preparations to assess synapse composition. While this supports the feasibility of using synaptosomes for proteomics-based comparisons, the resulting large datasets have not been very useful since they reflect a highly complex starting material containing an enormously heterogeneous population of synapses. Synapse heterogeneity is based largely on differences in connectivity, but currently there are no methodologies available that can be used to interrogate and analyze differences between identified synapse populations. Here we propose a novel and straightforward method based on the mammalian GFP reconstitution across synaptic partners technique that will enable proteomic comparisons between identified synapse populations in health and disease. We have assembled a team to test this using well- defined synaptic circuits. This approach will be a major step forward as it wll permit a rapid, large-scale assessment of particular synapse populations that can be isolated from complex circuits.