Li Binglun, Chavarha Mariya, Kobayashi Yuho, Yoshinaga Satoshi, Nakajima Kazunori, Lin Michael Z, Inoue Takafumi
Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda University, Tokyo 162-8480, Japan.
Department of Neurobiology, School of Medicine, Stanford University, Stanford, CA 94305-5090, USA.
iScience. 2020 Aug 21;23(8):101363. doi: 10.1016/j.isci.2020.101363. Epub 2020 Jul 12.
Recording the electrical activity of multiple neurons simultaneously would greatly facilitate studies on the function of neuronal circuits. The combination of the fast scanning by random-access multiphoton microscopy (RAMP) and the latest two-photon-compatible high-performance fluorescent genetically encoded voltage indicators (GEVIs) has enabled action potential detection in deep layers in in vivo brain. However, neuron connectivity analysis on optically recorded action potentials from multiple neurons in brain tissue has yet to be achieved. With high expression of a two-photon-compatible GEVI, ASAP3, via in utero electroporation and RAMP, we achieved voltage recording of spontaneous activities from multiple neurons in brain slice. We provide evidence for the developmental changes in intralaminar horizontal connections in somatosensory cortex layer 2/3 with a greater sensitivity than calcium imaging. This method thus enables investigation of neuronal network connectivity at the cellular resolution in brain tissue.
同时记录多个神经元的电活动将极大地促进对神经元回路功能的研究。随机存取多光子显微镜（RAMP）的快速扫描与最新的双光子兼容高性能荧光基因编码电压指示器（GEVIs）相结合，使得在活体大脑深层检测动作电位成为可能。然而，对脑组织中多个神经元光学记录的动作电位进行神经元连接性分析尚未实现。通过子宫内电穿孔和RAMP实现双光子兼容GEVI ASAP3的高表达，我们实现了脑片中多个神经元自发活动的电压记录。我们提供了体感皮层第2/3层层内水平连接发育变化的证据，其灵敏度高于钙成像。因此，该方法能够在细胞分辨率水平上研究脑组织中的神经网络连接性。
