title: Genetically encoded bioluminescent voltage indicator for multi-purpose use in wide range of bioimaging.
abstract: We report development of the first genetically encoded bioluminescent indicator for membrane voltage called LOTUS-V. Since it is bioluminescent, imaging LOTUS-V does not require external light illumination. This allows bidirectional optogenetic control of cellular activity triggered by Channelrhodopsin2 and Halorhodopsin during voltage imaging. The other advantage of LOTUS-V is the robustness of a signal-to-background ratio (SBR) wherever it expressed, even in the specimens where autofluorescence from environment severely interferes fluorescence imaging. Through imaging of moving cardiomyocyte aggregates, we demonstrated the advantages of LOTUS-V in long-term imaging are attributable to the absence of phototoxicity, and photobleaching in bioluminescent imaging, combined with the ratiometric aspect of LOTUS-V design. Collectively LOTUS-V extends the scope of excitable cell control and simultaneous voltage phenotyping, which should enable applications in bioscience, medicine and pharmacology previously not possible.
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Shigenori
authors.initials: S
authors.lastname: Inagaki
authors.affiliation: Department of Material Science, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi, Ishikawa 923-1292, Japan.|Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Tsutsui
authors.affiliation: Department of Biotechnology, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors.forename: Kazushi
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan.
authors.forename: Masakazu
authors.initials: M
authors.lastname: Agetsuma
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Arai
authors.affiliation: Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Jinno
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan.
authors.forename: Guirong
authors.initials: G
authors.lastname: Bai
authors.affiliation: BHF Centre for Regenerative Medicine, Division of Cardiovascular Medicine, West Wing Level 6, John Radcliffe Hospital, Oxford, OX3 9DU, UK.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Daniels
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan.|Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Okamura
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan.
authors.forename: Tomoki
authors.initials: T
authors.lastname: Matsuda
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan.|Department of Biotechnology, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.|The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan.
authors.forename: Takeharu
authors.initials: T
authors.lastname: Nagai
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D055724:Electrophysiological Phenomena; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism; D007700:Kinetics; D008163:Luminescent Measurements / Q000379:methods; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008958:Models, Molecular; D057054:Molecular Imaging / Q000379:methods; D062308:Optogenetics / Q000379:methods; D011487:Protein Conformation
chemical_list: D008164:Luminescent Proteins