Institute for Biological Sciences, National Research Council of Canada, 1200 Montreal Road, Ottawa, ON, K1A 0R6, Canada.
J Neural Eng. 2011 Jun;8(3):034002. doi: 10.1088/1741-2560/8/3/034002. Epub 2011 May 4.
Planar patch-clamp chip technology has been developed to enhance the assessment of novel compounds for therapeutic efficacy and safety. However, this technology has been limited to recording ion channels expressed in isolated suspended cells, making the study of ion channel function in synaptic transmission impractical. Recently, we developed single- and dual-recording site planar patch-clamp chips and demonstrated their capacity to record ion channel activity from neurons established in culture. Such capacity provides the opportunity to record from synaptically connected neurons cultured on-chip. In this study we reconstructed, on-chip, a simple synaptic circuit between cultured pre-synaptic visceral dorsal 4 neurons and post-synaptic left pedal dorsal 1 neurons isolated from the mollusk Lymnaea stagnalis. Here we report the first planar patch-clamp chip recordings of synaptic phenomena from these paired neurons and pharmacologically demonstrate the cholinergic nature of this synapse. We also report simultaneous dual-site recordings from paired neurons, and demonstrate dedicated cytoplasmic perfusion of individual neurons via on-chip subterranean microfluidics. This is the first application of planar patch-clamp technology to examine synaptic communication.
平面片钳芯片技术已经被开发出来，以增强对新型化合物的治疗效果和安全性的评估。然而，该技术仅限于记录在悬浮细胞中表达的离子通道，使得研究突触传递中的离子通道功能变得不切实际。最近，我们开发了单和双记录位点平面片钳芯片，并证明了它们从培养的神经元中记录离子通道活性的能力。这种能力为记录培养在芯片上的突触连接的神经元提供了机会。在这项研究中，我们在芯片上重建了来自腹侧 4 神经元的简单突触回路，该神经元是从软体动物田螺中分离出来的，而另一个神经元是左足背 1 神经元。在这里，我们报告了来自这些配对神经元的突触现象的首次平面片钳芯片记录，并通过药理学证明了这个突触的胆碱能性质。我们还报告了来自配对神经元的同时双位点记录，并通过芯片下的地下微流控技术证明了对单个神经元的专用细胞质灌注。这是平面片钳技术首次应用于研究突触通讯。
