Discovery of an F-actin–binding small molecule serving as a fluorescent probe and a scaffold for functional probes

Takagi, Takeru; Ueno, Takahiro; Ikawa, Keisuke; Asanuma, Daisuke; Nomura, Yusuke; Uno, Shin‐nosuke; Komatsu, Toshimitsu; Kamiya, Mako; Hanaoka, Kenjiro; Okimura, Chika; Iwadate, Yoshiaki; Hirose, Keikichi; Nagano, Tetsuo; Sugimura, Kaoru; Urano, Yasuteru

doi:10.1126/sciadv.abg8585

Cited by 14 publications

(9 citation statements)

References 38 publications

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“…45,46 Takagi et al recently developed an F-actin-binding dye, named HMRef, on the HeLa cell line. 47 Our observations also demonstrated the selective binding of CTC with the F-actin filament of both cell lines with high efficiency. Hence, it was confirmed giving a shred of strong evidence to us that synthesized CTC dye bind to F-actin with high accuracy.…”

Section: ■ Discussionsupporting

confidence: 63%

“…SCC-9 cells also have actin filaments and are used to study the various parameters of actin-related diseases . Hence, these cell lines are best to study the dye related to actin binding. , Takagi et al recently developed an F-actin-binding dye, named HMRef, on the HeLa cell line . Our observations also demonstrated the selective binding of CTC with the F-actin filament of both cell lines with high efficiency.…”

Section: Discussionsupporting

confidence: 62%

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Synthesis of First Coumarin Fluorescent Dye for Actin Visualization

et al. 2022

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Selective fluorescence imaging of actin protein hugely depends on the fluorescently labeled actin-binding domain (ABD). Thus, it is always a challenging task to image the actin protein (in vivo or in vitro) directly with an ABD-free system. To overcome the limitations of actin imaging without an ABD, we have designed a facile and cost-effective red fluorescent coumarin dye 7-hydroxy-4-methyl-8-(4-(2-oxo-2H-chromen-3-yl)thiazol-2-ylimino)methyl-2H-chromen-2-one (CTC) for actin binding. The selective binding of the dye was investigated using the gut and eye of the model organism Drosophila melanogaster and C2C12 and SCC-9 cell lines. Our result suggests two major advantages of CTC over the dyes presently used for imaging actin proteins. First, the dye can bind to actin efficiently without any secondary intermediate. Second, it is much more stable at room temperature and exhibits excellent photostability. To the best of our knowledge, this is the first fluorescent dye that can bind to the actin protein without employing any secondary intermediate/actin-binding domain. These findings could pave the way for many biologists and physicists to successfully employ the CTC dye for imaging and tracking actin proteins by fluorescence microscopy in various in vivo and in vitro systems.

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Section: ■ Discussionsupporting

confidence: 63%

Section: Discussionsupporting

confidence: 62%

Synthesis of First Coumarin Fluorescent Dye for Actin Visualization

et al. 2022

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“…5 A and Movie S10 ). F-actin fluorescence, which discloses actomyosin cables, was visualized with HMRef ( 41 ) in a living cell sheet. The rectangle in 13.5 min in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…HMRef, a novel probe for F-actin developed by Ueno and colleagues ( 41 ), was dissolved at 1 mM in DMSO and then diluted 4,000-fold with culture medium. This HMRef medium was then added to the chamber, to the bottom of which the cell sheets had adhered, immediately after removal of the culture medium.…”

Section: Methodsmentioning

confidence: 99%

Leading-edge elongation by follower cell interruption in advancing epithelial cell sheets

Okimura

Iwanaga

Sakurai

et al. 2022

Proc. Natl. Acad. Sci. U.S.A.

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Significance Collective cell migration is an important phenomenon in the developmental process of living organisms and even in mature individuals. The cells in the collective are accurately classified into leader cells, which lead the collective at the leading edge, and follower cells. We found that in the collective migration of keratocytes, the epidermal cells of fish responsible for wound repair, a mechanical interaction between two leader cells and a single follower cell can forcibly break the connection between the two leader cells and transform the follower cell into a leader cell.

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“…HMRef, an F-actin staining reagent, was a kind gift from Prof. Y. Urano (the University of Tokyo, Japan). 13 Other reagents, including salts and culture media, were obtained from Sigma-Aldrich and Fujifilm Wako unless otherwise specified.…”

Section: Methodsmentioning

confidence: 99%

A noncanonical endocytic pathway is involved in the internalization of 3 μm polystyrene beads into HeLa cells

et al. 2022

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Discovery of an F-actin–binding small molecule serving as a fluorescent probe and a scaffold for functional probes

Abstract: We found an F-actin–binding small molecule serving as a fluorescent probe and a scaffold for functional probes.

Cited by 14 publications

References 38 publications

Synthesis of First Coumarin Fluorescent Dye for Actin Visualization

Synthesis of First Coumarin Fluorescent Dye for Actin Visualization

Leading-edge elongation by follower cell interruption in advancing epithelial cell sheets

A noncanonical endocytic pathway is involved in the internalization of 3 μm polystyrene beads into HeLa cells

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