Rational Engineering of XCaMPs, a Multicolor GECI Suite for In Vivo Imaging of Complex Brain Circuit Dynamics

Masatoshi Inoue; Atsuya Takeuchi; Satoshi Manita; Shin-Ichiro Horigane; Masayuki Sakamoto; Ryosuke Kawakami; Kazushi Yamaguchi; Kouhei Otomo; Hiroyuki Yokoyama; Ryang Kim; Tatsushi Yokoyama; Sayaka Takemoto-Kimura; Manabu Abe; Michiko Okamura; Yayoi Kondo; Sean Quirin; Charu Ramakrishnan; Takeshi Imamura; Kenji Sakimura; Tomomi Nemoto; Masanobu Kano; Hajime Fujii; Karl Deisseroth; Kazuo Kitamura; Haruhiko Bito

doi:10.1016/j.cell.2019.04.007

Rational Engineering of XCaMPs, a Multicolor GECI Suite for In Vivo Imaging of Complex Brain Circuit Dynamics

Cell. 2019 May 16;177(5):1346-1360.e24. doi: 10.1016/j.cell.2019.04.007. Epub 2019 May 9.

Authors

Masatoshi Inoue¹, Atsuya Takeuchi², Satoshi Manita³, Shin-Ichiro Horigane⁴, Masayuki Sakamoto⁵, Ryosuke Kawakami⁶, Kazushi Yamaguchi⁷, Kouhei Otomo⁷, Hiroyuki Yokoyama⁸, Ryang Kim⁵, Tatsushi Yokoyama⁵, Sayaka Takemoto-Kimura⁹, Manabu Abe¹⁰, Michiko Okamura⁵, Yayoi Kondo⁵, Sean Quirin¹¹, Charu Ramakrishnan¹¹, Takeshi Imamura¹², Kenji Sakimura¹⁰, Tomomi Nemoto¹³, Masanobu Kano¹⁴, Hajime Fujii⁵, Karl Deisseroth¹¹, Kazuo Kitamura¹⁵, Haruhiko Bito¹⁶

Affiliations

¹ Department of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan; Department of Bioengineering, Stanford University School of Medicine, Stanford, CA 94305, USA.
² Department of Neurophysiology, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
³ Department of Neurophysiology, Faculty of Medicine, University of Yamanashi, Chuo, Yamanashi 409-3898, Japan.
⁴ Department of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan; Department of Neuroscience I, Research Institute of Environmental Medicine, Nagoya University, Nagoya, Aichi 464-8601, Japan; Department of Molecular/Cellular Neuroscience, Nagoya University Graduate School of Medicine, Nagoya, Aichi 466-8550, Japan.
⁵ Department of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
⁶ Laboratory of Molecular and Cellular Biophysics, Research Institute for Electronic Science, Hokkaido University, Sapporo, Hokkaido 001-0020, Japan; Department of Molecular Medicine for Pathogenesis, Ehime University Graduate School of Medicine, Toon, Ehime 791-0295, Japan.
⁷ Laboratory of Molecular and Cellular Biophysics, Research Institute for Electronic Science, Hokkaido University, Sapporo, Hokkaido 001-0020, Japan.
⁸ New Industry Creation Hatchery Center (NICHe), Tohoku University, Sendai, Miyagi 980-8579, Japan.
⁹ Department of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan; Department of Neuroscience I, Research Institute of Environmental Medicine, Nagoya University, Nagoya, Aichi 464-8601, Japan; Department of Molecular/Cellular Neuroscience, Nagoya University Graduate School of Medicine, Nagoya, Aichi 466-8550, Japan; Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency, Saitama, Japan.
¹⁰ Department of Cellular Neurobiology, Brain Research Institute, Niigata University, Niigata, Niigata 951-8585, Japan.
¹¹ Department of Bioengineering, Stanford University School of Medicine, Stanford, CA 94305, USA.
¹² Department of Molecular Medicine for Pathogenesis, Ehime University Graduate School of Medicine, Toon, Ehime 791-0295, Japan.
¹³ Laboratory of Molecular and Cellular Biophysics, Research Institute for Electronic Science, Hokkaido University, Sapporo, Hokkaido 001-0020, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Agency for Medical Research and Development, Tokyo, Japan.
¹⁴ Department of Neurophysiology, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan; International Research Center for Neurointelligence, The University of Tokyo, Tokyo, Japan.
¹⁵ Department of Neurophysiology, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan; Department of Neurophysiology, Faculty of Medicine, University of Yamanashi, Chuo, Yamanashi 409-3898, Japan.
¹⁶ Department of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan; International Research Center for Neurointelligence, The University of Tokyo, Tokyo, Japan. Electronic address: hbito@m.u-tokyo.ac.jp.

PMID: 31080068
DOI: 10.1016/j.cell.2019.04.007

Abstract

To decipher dynamic brain information processing, current genetically encoded calcium indicators (GECIs) are limited in single action potential (AP) detection speed, combinatorial spectral compatibility, and two-photon imaging depth. To address this, here, we rationally engineered a next-generation quadricolor GECI suite, XCaMPs. Single AP detection was achieved within 3-10 ms of spike onset, enabling measurements of fast-spike trains in parvalbumin (PV)-positive interneurons in the barrel cortex in vivo and recording three distinct (two inhibitory and one excitatory) ensembles during pre-motion activity in freely moving mice. In vivo paired recording of pre- and postsynaptic firing revealed spatiotemporal constraints of dendritic inhibition in layer 1 in vivo, between axons of somatostatin (SST)-positive interneurons and apical tufts dendrites of excitatory pyramidal neurons. Finally, non-invasive, subcortical imaging using red XCaMP-R uncovered somatosensation-evoked persistent activity in hippocampal CA1 neurons. Thus, the XCaMPs offer a critical enhancement of solution space in studies of complex neuronal circuit dynamics. VIDEO ABSTRACT.

Keywords: PV recording; XCaMP; ckkap sequence; genetically encoded calcium indicators; multiplex imaging; non-invasive hippocampal recording; paired pre- and post-synapse recording; two-photon Ca(2+) imaging.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Action Potentials / physiology*
Animals
Axons / metabolism*
Cerebral Cortex / cytology
Cerebral Cortex / metabolism*
Female
Hippocampus / cytology
Hippocampus / metabolism*
Interneurons / cytology
Interneurons / metabolism*
Mice
Mice, Transgenic
Pyramidal Cells / cytology
Pyramidal Cells / metabolism*
Rats
Rats, Sprague-Dawley

Grants and funding

HHMI/Howard Hughes Medical Institute/United States