Detection of LacZ-Positive Cells in Living Tissue with Single-Cell Resolution

Tomohiro Doura, Mako Kamiya, Fumiaki Obata, Yoshifumi Yamaguchi, Takeshi Y. Hiyama, Takashi Matsuda, Akiyoshi Fukamizu, Masaharu Noda, Masayuki Miura, Yasuteru Urano

Research output: Contribution to journalArticle

21 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)9620-9624
Number of pages5
JournalAngewandte Chemie - International Edition
Volume55
Issue number33
DOIs
Publication statusPublished - Jan 1 2016
Externally publishedYes

Fingerprint

Galactosidases
Cells
Tissue
Fluorescent Dyes
Gene expression
Labeling
Escherichia coli
Neurons
Enzymes
Genes
Fluorescence
Chemical activation
Ligands
Proteins
Substrates
single cell proteins

Keywords

  • fluorescent probes
  • lacZ
  • quinone methide intermediates
  • single-cell resolution
  • β-galactosidase

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)

Cite this

Detection of LacZ-Positive Cells in Living Tissue with Single-Cell Resolution. / Doura, Tomohiro; Kamiya, Mako; Obata, Fumiaki; Yamaguchi, Yoshifumi; Hiyama, Takeshi Y.; Matsuda, Takashi; Fukamizu, Akiyoshi; Noda, Masaharu; Miura, Masayuki; Urano, Yasuteru.

In: Angewandte Chemie - International Edition, Vol. 55, No. 33, 01.01.2016, p. 9620-9624.

Research output: Contribution to journalArticle

Doura, T, Kamiya, M, Obata, F, Yamaguchi, Y, Hiyama, TY, Matsuda, T, Fukamizu, A, Noda, M, Miura, M & Urano, Y 2016, 'Detection of LacZ-Positive Cells in Living Tissue with Single-Cell Resolution', Angewandte Chemie - International Edition, vol. 55, no. 33, pp. 9620-9624. https://doi.org/10.1002/anie.201603328
Doura, Tomohiro ; Kamiya, Mako ; Obata, Fumiaki ; Yamaguchi, Yoshifumi ; Hiyama, Takeshi Y. ; Matsuda, Takashi ; Fukamizu, Akiyoshi ; Noda, Masaharu ; Miura, Masayuki ; Urano, Yasuteru. / Detection of LacZ-Positive Cells in Living Tissue with Single-Cell Resolution. In: Angewandte Chemie - International Edition. 2016 ; Vol. 55, No. 33. pp. 9620-9624.
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