TY - JOUR
T1 - Two-Photon Voltage Imaging of Spontaneous Activity from Multiple Neurons Reveals Network Activity in Brain Tissue
AU - Li, Binglun
AU - Chavarha, Mariya
AU - Kobayashi, Yuho
AU - Yoshinaga, Satoshi
AU - Nakajima, Kazunori
AU - Lin, Michael Z.
AU - Inoue, Takafumi
N1 - Funding Information:
This work was supported by Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS; JP16H06482, K.N.; 23300121, T.I.), Private University Research Branding Project (MEXT, Japan, T.I.), and Waseda University Grants for Special Research Projects (Waseda University, Japan, T.I.). All the authors had full access to all the data in this study and take responsibility for the integrity of the data and the accuracy of the data analysis. T.I. and B.L. conceived and designed the research; B.L. preformed the research and analyzed the data; Y.K. contributed in molecular biology and microscopy; B.L. learned the IUE protocol from S.Y. and K.N. at K.N.’s laboratory in Keio University School of Medicine; M.C. and M.Z.L. created and provided ASAP3; B.L and T.I. drafted the manuscript; M.C. and M.Z.L. provided critical revision of the manuscript; T.I. obtained funding. The authors declare no competing interests.
Funding Information:
This work was supported by Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS; JP16H06482 , K.N.; 23300121 , T.I.), Private University Research Branding Project ( MEXT , Japan, T.I.), and Waseda University Grants for Special Research Projects (Waseda University, Japan, T.I.).
Publisher Copyright:
© 2020 The Author(s)
PY - 2020/8/21
Y1 - 2020/8/21
N2 - 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.
AB - 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.
KW - Cellular Neuroscience
KW - Developmental Neuroscience
KW - Optical Imaging
KW - Techniques in Neuroscience
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U2 - 10.1016/j.isci.2020.101363
DO - 10.1016/j.isci.2020.101363
M3 - Article
AN - SCOPUS:85088398745
SN - 2589-0042
VL - 23
JO - iScience
JF - iScience
IS - 8
M1 - 101363
ER -