Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, Japan.
Center for Information and Neural Networks, National Institute of Information and Communications Technology, Suita City, Osaka, 565-0871, Japan.
STAR Protoc. 2020 Oct 6;1(3):100121. doi: 10.1016/j.xpro.2020.100121. eCollection 2020 Dec 18.
Most excitatory inputs arrive at dendritic spines in a postsynaptic neuron. To understand dendritic information processing, it is critical to scrutinize the spatiotemporal dynamics of synaptic inputs along dendrites. This protocol combines spinning-disk confocal imaging with whole-cell patch-clamp recording to perform wide-field, high-speed optical recording of synaptic inputs in a neuron loaded with a calcium indicator in cultured networks. Our protocol enables simultaneous detection of synaptic inputs as calcium signals from hundreds of spines in multiple dendritic branches. For complete details on the use and execution of this protocol, please refer to Takahashi et al. (2012, 2016), Kobayashi et al. (2019), and Ishikawa and Ikegaya (2020).
大多数兴奋性输入到达神经元的树突棘。为了理解树突信息处理，关键是仔细研究沿着树突的突触输入的时空动态。本方案结合旋转盘共聚焦成像和全细胞膜片钳记录，在培养网络中用钙指示剂加载的神经元中进行突触输入的宽场、高速光学记录。我们的方案能够同时检测来自多个树突分支中数百个树突棘的作为钙信号的突触输入。有关此方案的使用和执行的完整详细信息，请参阅 Takahashi 等人（2012 年，2016 年）、Kobayashi 等人（2019 年）和 Ishikawa 和 Ikegaya（2020 年）。
