Pons-Bennaceur Alexandre, Lozovaya Natalia
INSERM U901, Mediterranean Institute of Neurobiology (INMED), Aix-Marseille University, Marseille, France.
Neurochlore c/o Mediterranean Institute of Neurobiology (INMED), INSERM U901, Parc Scientifique de Luminy, BP13, 13273, Marseille Cedex 09, France.
Methods Mol Biol. 2017;1677:231-239. doi: 10.1007/978-1-4939-7321-7_12.
Analysis of electrophysiological properties of NMDARs and NMDAR-mediated synaptic transmission in identified neurons and synapses in brain slices is a major step in understanding their function in normal and pathological neuronal brain networks. In many central synapses excitatory postsynaptic currents (EPSCs) are mediated by excitatory neurotransmitter glutamate that activates colocalized AMPAR and NMDAR generating a complex EPSC. Here, we describe the methods commonly used in brain slices to study the electrophysiological properties of NMDAR-mediated component of spontaneous or evoked EPSCs by extracellular stimulation or by stimulating synaptically connected neurons. This approach is based on whole-cell patch-clamp recordings, pharmacological tools, and the analysis of the difference in temporal parameters between the AMPA and NMDA receptors. It allows pinpointing of the basic functional properties of NMDARs that are specific to identified brain regions, neurons, and synapses of wild-type or genetically manipulated mice.
分析脑片中特定神经元和突触中NMDARs的电生理特性以及NMDAR介导的突触传递,是理解它们在正常和病理性神经元脑网络中功能的重要一步。在许多中枢突触中,兴奋性突触后电流(EPSCs)由兴奋性神经递质谷氨酸介导,谷氨酸激活共定位的AMPAR和NMDAR,产生复杂的EPSC。在这里,我们描述了在脑片中常用的方法,通过细胞外刺激或刺激突触连接的神经元,来研究自发或诱发EPSCs中NMDAR介导成分的电生理特性。这种方法基于全细胞膜片钳记录、药理学工具以及对AMPA和NMDA受体之间时间参数差异的分析。它能够精确确定野生型或基因操作小鼠特定脑区、神经元和突触中NMDARs的基本功能特性。