SPECIFIC PROTEOME OF MAMMALIAN CORTEX INHIBITORY & EXCITATORY SYNAPSES

哺乳动物皮层抑制的特定蛋白质组

• 批准号: 8169160
• 负责人: NATHANIEL HEINTZ
• 金额: $ 5.24万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169160
• 关键词: Bacterial Artificial Chromosomes; Biology; Cells; Cerebellum; Collaborations; Complement; Computer Retrieval of Information on Scientific Projects Database; Dissection; Excitatory Synapse; Fiber; Funding; GABA Receptor; Grant; Hares; Inhibitory Synapse; Institution; Manuscripts; Modification; Mus; Population; Proteins; Proteome; Proteomics; Publishing; Purkinje Cells; Research; Research Personnel; Resources; Source; Synapses; United States National Institutes of Health; Universities; Work; in vivo; novel

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Four years ago, in collaboration with Nat Heintz (Rockefeller University), we initiated the study of the protein complement present at excitatory synapses in Purkinje cells. We used the Bacterial Artificial Chromosome (BAC) modification strategy to target the specific in vivo expression of GFP-fused GRID 2 to Purkinje cell's excitatory synapses. We performed dissections of mouse cerebella, and purified synapses bearing GFP-GRID2. Although challenging, our approach proved successful, as we isolated synapses and analyzed low-femtomol levels of proteins. During this last year, we continued our mass spectrometric analyses and identified ~70 synaptic proteins, confirming known excitatory proteins, the absence of inhibitory proteins, and identifying novel signatures of excitatory synapses. We have published a manuscript describing this work (F. Selimi, I. Cristea, E. Heller, B.T. Chait, N. Heintz "Proteomic studies of a single CNS synapse type: the parallel fiber/Purkinje cell synapse" PLoS Biology, 2009 Apr 14;7(4):e83). Using a similar approach to the one described above, we hare currently studying the protein composition of inhibitory synapses by isolating GABA receptors from specific cell populations in the Cortex.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 四年前，我们与Nat Heintz（洛克菲勒大学）合作，开始了对浦肯野细胞兴奋性突触中存在的蛋白质补体的研究。我们采用细菌人工染色体（BAC）修饰策略，在浦肯野细胞的兴奋性突触中特异性表达GFP融合的GRID 2。我们进行了小鼠小脑的解剖，并纯化了带有GFP-GRID 2的突触。虽然具有挑战性，但我们的方法被证明是成功的，因为我们分离了突触并分析了低飞摩尔水平的蛋白质。在过去的一年里，我们继续进行质谱分析，鉴定了约70种突触蛋白，确认了已知的兴奋性蛋白，没有抑制性蛋白，并鉴定了兴奋性突触的新特征。我们已经出版了一份手稿描述这项工作（F。塞利米岛Cristea，E. B.T.海勒Chait，N. Heintz“Proteomic studies of a single CNS synapse type：the parallel fiber/Purkinje cell synapse”PLoS Biology，2009 Apr 14;7（4）：e83）。 使用与上述方法类似的方法，我们目前正在研究抑制性突触的蛋白质组成，方法是从皮层的特定细胞群中分离GABA受体。