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制备突触神经小体以研究小鼠大脑皮层中的突触

Preparation of Synaptoneurosomes to Study the Synapse in the Murine Cerebral Cortex.

作者信息

Diaz Ariel, Torre Enrique, Yepes Manuel

机构信息

Division of Neuropharmacology and Neurologic Diseases, Yerkes National Primate Research Center, Atlanta, GA; USA.

Department of Neurology, Emory University, Atlanta, GA; USA.

出版信息

Bio Protoc. 2021 Jan 20;11(2):e3896. doi: 10.21769/BioProtoc.3896.

Abstract

The synapse is a complex structure where the transmission of information takes place. Synaptic dysfunction is one of the earliest pathophysiological events in several diseases, such as traumatic brain injury, cerebral ischemia, and neurodegenerative diseases. Thus, a methodology to study synaptic structure and function is crucial for the development of potential strategies for the treatment of many neurological diseases. Synaptoneurosomes (SNs) are structures assembled by the sealed presynaptic bouton and the attached post-synaptic density. Despite the fact that for a long time it has been recognized that SNs are a powerful tool to study synaptic function, composition, and structure, its use has been limited by the requirement of relatively large amounts of material to successfully isolate them. Here we describe a three-step centrifugation procedure performed under hypotonic conditions to isolate SNs from small volumes of the cerebral cortex.

摘要

突触是信息传递发生的复杂结构。突触功能障碍是多种疾病(如创伤性脑损伤、脑缺血和神经退行性疾病)中最早出现的病理生理事件之一。因此,研究突触结构和功能的方法对于开发许多神经疾病的潜在治疗策略至关重要。突触神经小体(SNs)是由密封的突触前终扣和附着的突触后致密物组装而成的结构。尽管长期以来人们一直认识到SNs是研究突触功能、组成和结构的有力工具,但其应用受到成功分离所需相对大量材料的限制。在此,我们描述了一种在低渗条件下进行的三步离心程序,用于从小体积的大脑皮层中分离SNs。

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