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Calponin 3 regulates stress fiber formation in dermal fibroblasts during wound healing

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Abstract

Skin wound healing is an intricate process involving various cell types and molecules. In granulation tissue, fibroblasts proliferate and differentiate into myofibroblasts and generate mechanical tension for wound closure and contraction. Actin stress fibers formed in these cells, especially those containing α-smooth muscle actin (α-SMA), are the central machinery for contractile force generation. In the present study, calponin 3 (CNN3), which has a phosphorylation-dependent actin-binding property, was identified in the molecular mechanism underlying stress fiber formation. CNN3 was expressed by fibroblasts/myofibroblasts in the proliferation phase of wound healing, and was associated with α-SMA in stress fibers formed by cultured dermal fibroblasts. CNN3 expression was post-transcriptionally regulated by tension, as demonstrated by disruption of actin filament organization under floating culture or blebbistatin treatment. CNN3 knockdown in primary fibroblasts impaired stress fiber formation, resulting in a phenotype of decreased cellular dynamics such as cell motility and contractile ability. These findings indicate that CNN3 participates in actin stress fiber remodeling, which is required for cell motility and contraction of dermal fibroblasts in the wound healing process.

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Abbreviations

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

Fetal bovine serum

PBS:

Phosphate-buffered saline

CNN3:

Calponin 3

α-SMA:

α-Smooth muscle actin

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Acknowledgments

We thank Dr Hiroyuki Miyoshi (RIKEN BRC) for providing lentivirus vectors. This work was supported in part by Grants-in-Aid for Scientific Research (B) (23390081), (C) (24591618) and the Global Centers of Excellence (GCOE) Program of Osaka University from the Japan Society for the Promotion of Science (JSPS).

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The authors have no conflict of interest to disclose.

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Authors and Affiliations

  1. Department of Molecular Medicine, Osaka Medical Center and Research Institute for Maternal and Child Health, 840 Murodo-cho, Izumi, Osaka, 594-1101, Japan

    Etsuko Daimon, Yukinao Shibukawa & Yoshinao Wada

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  1. Etsuko Daimon
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Correspondence to Yoshinao Wada.

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Daimon, E., Shibukawa, Y. & Wada, Y. Calponin 3 regulates stress fiber formation in dermal fibroblasts during wound healing. Arch Dermatol Res 305, 571–584 (2013). https://doi.org/10.1007/s00403-013-1343-8

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