Mass synaptometry: High-dimensional multi parametric assay for single synapses
Mass synaptometry: High-dimensional multi parametric assay for single synapses
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DOI：
10.1016/j.jneumeth.2018.11.008
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发表时间：
2019-01-15
影响因子：
3
通讯作者：
Montine, Thomas J.
中科院分区：
文献类型：
--
作者：
Gajera, Chandresh R.;Fernandez, Rosemary;Montine, Thomas J.
背景：突触改变，尤其是突触前改变，是神经退行性疾病的基本特征，与认知功能下降密切相关。新方法：我们报道了一种用于对单个人类突触小体进行高维分析的方法，丰富了大脑中的神经末梢。这种方法是从免疫和血细胞的单细胞分析中常用的飞行时间质谱仪(CyTOF)的细胞学方法改编而来的。结果：在这里，我们通过优化突触体准备、生成“SynTOF面板”、重新校准采集设置和应用计算分析，克服了单一突触分析的挑战。通过对390,000个单个突触体的分析，我们还通过表征路易体病、阿尔茨海默病和正常大脑中突触多样性的变化，提供了原理验证的证据。与现有方法(S)相比：目前用于研究人类突触的成像方法能够分析有限数量的突触，而传统的流式细胞仪技术通常被限制在6个参数以下。我们的方法可以从数万个单个突触中同时检测34个参数。结论：我们应用质量突触测量法同时分析了13个人脑样本中39万多个突触的34个参数。这一新方法揭示了突触表型的区域性和疾病特异性变化，包括用路易体病和阿尔茨海默病纹状体多巴胺能突触分子组成的预期变化来验证这一方法。质量突触测量术使单个突触终末的高度平行的分子图谱成为可能。
Background: Synaptic alterations, especially presynaptic changes, are cardinal features of neurodegenerative diseases and strongly correlate with cognitive decline.New method: We report "Mass Synaptometry" for the high-dimensional analysis of individual human synaptosomes, enriched nerve terminals from brain. This method was adapted from cytometry by time-of-flight mass spectrometry (CyTOF), which is commonly used for single-cell analysis of immune and blood cells.Result: Here we overcome challenges for single synapse analysis by optimizing synaptosome preparations, generating a 'SynTOF panel,' recalibrating acquisition settings, and applying computational analyses. Through the analysis of 390,000 individual synaptosomes, we also provide proof-of principle validation by characterizing changes in synaptic diversity in Lewy Body Disease (LBD), Alzheimer's disease and normal brain.Comparison with existing method(s): Current imaging methods to study synapses in humans are capable of analyzing a limited number of synapses, and conventional flow cytometric techniques are typically restricted to fewer than 6 parameters. Our method allows for the simultaneous detection of 34 parameters from tens of thousands of individual synapses.Conclusion: We applied Mass Synaptometry to analyze 34 parameters simultaneously on more than 390,000 synaptosomes from 13 human brain samples. This new approach revealed regional and disease-specific changes in synaptic phenotypes, including validation of this method with the expected changes in the molecular composition of striatal dopaminergic synapses in Lewy body disease and Alzheimer's disease. Mass synaptometry enables highly parallel molecular profiling of individual synaptic terminals.