Debanne Dominique, Boudkkazi Sami, Campanac Emilie, Cudmore Robert H, Giraud Pierre, Fronzaroli-Molinieres Laure, Carlier Edmond, Caillard Olivier
Institut National de la Santé et de la Recherche Médicale U641, Marseille, France.
Nat Protoc. 2008;3(10):1559-68. doi: 10.1038/nprot.2008.147.
Analysis of synaptic transmission, synaptic plasticity, axonal processing, synaptic timing or electrical coupling requires the simultaneous recording of both the pre- and postsynaptic compartments. Paired-recording technique of monosynaptically connected neurons is also an appropriate technique to probe the function of small molecules (calcium buffers, peptides or small proteins) at presynaptic terminals that are too small to allow direct whole-cell patch-clamp recording. We describe here a protocol for obtaining, in acute and cultured slices, synaptically connected pairs of cortical and hippocampal neurons, with a reasonably high probability. The protocol includes four main stages (acute/cultured slice preparation, visualization, recording and analysis) and can be completed in approximately 4 h.
对突触传递、突触可塑性、轴突加工、突触时间或电耦合进行分析,需要同时记录突触前和突触后部分。单突触连接神经元的配对记录技术也是一种合适的技术,用于探测突触前终末太小而无法进行直接全细胞膜片钳记录的小分子(钙缓冲剂、肽或小蛋白质)的功能。我们在此描述一种方案,该方案能够以相当高的概率在急性切片和培养切片中获得皮质和海马神经元的突触连接对。该方案包括四个主要阶段(急性/培养切片制备、可视化、记录和分析),大约4小时即可完成。