Postupna Nadia O, Latimer Caitlin S, Keene C Dirk, Montine Kathleen S, Montine Thomas J, Darvas Martin
Department of Pathology, University of Washington, Seattle WA, USA.
Department of Pathology, Stanford University, Stanford CA, USA.
Neuromethods. 2018;141:297-310. doi: 10.1007/978-1-4939-8739-9_17. Epub 2018 Sep 22.
Neurodegenerative diseases, the most common among them Alzheimer's disease (AD) and Lewy body disease (LBD), are a group of progressive incurable illnesses. In both AD and LBD, abundant evidence points to the synapse as the critical and early focus of pathological changes. Here we present a method for the isolation and flow cytometric analysis of synaptosomes prepared from postmortem human brain tissue, which we also applied to animal models, including mice and nonhuman primates. The use of flow cytometry for analysis allows for relatively fast and efficient examination of thousands of synaptosome particles in a matter of minutes, and also makes it possible to use crude, rather than purified, synaptosomal preparation, thus conserving tissue resources. We have applied this method to study synaptic alteration in several brain regions in human research participants and animal models.
神经退行性疾病,其中最常见的是阿尔茨海默病(AD)和路易体病(LBD),是一组进行性的不治之症。在AD和LBD中,大量证据表明突触是病理变化的关键和早期焦点。在此,我们介绍一种从死后人类脑组织制备的突触体的分离和流式细胞术分析方法,该方法我们也应用于动物模型,包括小鼠和非人类灵长类动物。使用流式细胞术进行分析能够在几分钟内相对快速且高效地检测数千个突触体颗粒,并且还使得使用粗制而非纯化的突触体制备成为可能,从而节省组织资源。我们已将此方法应用于研究人类研究参与者和动物模型中几个脑区的突触改变。