title: Detection of LacZ-Positive Cells in Living Tissue with Single-Cell Resolution.
abstract: The LacZ gene, which encodes Escherichia coli β-galactosidase, is widely used as a marker for cells with targeted gene expression or disruption. However, it has been difficult to detect lacZ-positive cells in living organisms or tissues at single-cell resolution, limiting the utility of existing lacZ reporters. Herein we present a newly developed fluorogenic β-galactosidase substrate suitable for labeling live cells in culture, as well as in living tissues. This precisely functionalized fluorescent probe exhibited dramatic activation of fluorescence upon reaction with the enzyme, remained inside cells by anchoring itself to intracellular proteins, and provided single-cell resolution. Neurons labeled with this probe preserved spontaneous firing, which was enhanced by application of ligands of receptors expressed in the cells, suggesting that this probe would be applicable to investigate functions of targeted cells in living tissues and organisms.
authors.affiliation: Graduate School of Medicine, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Doura
authors.affiliation: Graduate School of Medicine, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan. mkamiya@m.u-tokyo.ac.jp.|PRESTO, Japan, Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama, 332-0012, Japan. mkamiya@m.u-tokyo.ac.jp.
authors.forename: Mako
authors.initials: M
authors.lastname: Kamiya
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Obata
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.|PRESTO, Japan, Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama, 332-0012, Japan.
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Yamaguchi
authors.affiliation: Division of Molecular Neurobiology, National Institute for Basic Biology, 5-1 Higashiyama, Myodaiji-cho, Okazaki, Aichi, 444-8787, Japan.|School of Life Science, The Graduate University for Advanced Studies, 5-1 Higashiyama, Myodaiji-cho, Okazaki, Aichi, 444-8787, Japan.
authors.forename: Takeshi Y
authors.initials: TY
authors.lastname: Hiyama
authors.affiliation: School of Life Science, The Graduate University for Advanced Studies, 5-1 Higashiyama, Myodaiji-cho, Okazaki, Aichi, 444-8787, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Matsuda
authors.affiliation: Life Science Center, Tsukuba Advanced Research Alliance, Tsukuba, Ibaraki, 305-8577, Japan.|Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, 305-8577, Japan.
authors.forename: Akiyoshi
authors.initials: A
authors.lastname: Fukamizu
authors.affiliation: Division of Molecular Neurobiology, National Institute for Basic Biology, 5-1 Higashiyama, Myodaiji-cho, Okazaki, Aichi, 444-8787, Japan.|School of Life Science, The Graduate University for Advanced Studies, 5-1 Higashiyama, Myodaiji-cho, Okazaki, Aichi, 444-8787, Japan.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Noda
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.|CREST, Japan, Agency for Medical Research and Development, 1-7-1 Otemachi, Chiyoda-ku, Tokyo, 100-0004, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Miura
authors.affiliation: Graduate School of Medicine, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan. uranokun@m.u-tokyo.ac.jp.|Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan. uranokun@m.u-tokyo.ac.jp.|CREST, Japan, Agency for Medical Research and Development, 1-7-1 Otemachi, Chiyoda-ku, Tokyo, 100-0004, Japan. uranokun@m.u-tokyo.ac.jp.
authors.forename: Yasuteru
authors.initials: Y
authors.lastname: Urano
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004331:Drosophila melanogaster / Q000166:cytology*; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D007763:Lac Operon; D015394:Molecular Structure; D059010:Single-Cell Analysis; D001616:beta-Galactosidase / Q000737:chemistry* / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D001616:beta-Galactosidase
keywords: fluorescent probes; lacZ; quinone methide intermediates; single-cell resolution; β-galactosidase