The connexin mimetic peptide Gap27 and Cx43-knockdown reveal differential roles for Connexin43 in wound closure events in skin model systems

Faniku, C., O'Shaughnessy, E., Lorraine, C., Johnstone, S. R., Graham, A., Greenhough, S. and Martin, P. E.M. (2018) The connexin mimetic peptide Gap27 and Cx43-knockdown reveal differential roles for Connexin43 in wound closure events in skin model systems. International Journal of Molecular Sciences, 19(2), 604. (doi: 10.3390/ijms19020604) (PMID:29463027)

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Abstract

In the epidermis, remodelling of Connexin43 is a key event in wound closure. However, controversy between the role of connexin channel and non-channel functions exist. We compared the impact of SiRNA targeted to Connexin43 and the connexin mimetic peptide Gap27 on scrape wound closure rates and hemichannel signalling in adult keratinocytes (AK) and fibroblasts sourced from juvenile foreskin (JFF), human neonatal fibroblasts (HNDF) and adult dermal tissue (ADF). The impact of these agents, following 24 h exposure, on (encoding Connexin43), and gene expression, and Connexin43 and pSmad3 protein expression levels, were examined by qPCR and Western Blot respectively. In all cell types Gap27 (100-100 μM) attenuated hemichannel activity. In AK and JFF cells, Gap27 (100 nM-100 μM) enhanced scrape wound closure rates by ~50% but did not influence movement in HNDF or ADF cells. In both JF and AK cells, exposure to Gap27 for 24 h reduced the level of Cx43 protein expression but did not affect the level in ADF and HNDF cells. Connexin43-SiRNA enhanced scrape wound closure in all the cell types under investigation. In HDNF and ADF, Connexin43-SiRNA enhanced cell proliferation rates, with enhanced proliferation also observed following exposure of HDNF to Gap27. By contrast, in JFF and AK cells no changes in proliferation occurred. In JFF cells, Connexin43-SiRNA enhanced levels and in JFF and ADF cells both Connexin43-SiRNA and Gap27 enhanced pSmad3 protein expression levels. We conclude that Connexin43 signalling plays an important role in cell migration in keratinocytes and foreskin derived fibroblasts, however, different pathways are evoked and in dermal derived adult and neonatal fibroblasts, inhibition of Connexin43 signalling plays a more significant role in regulating cell proliferation than cell migration.

Item Type:Articles
Additional Information:Chrysovalantou Faniku and Claire Lorraine were funded by GCU studentships and Erin O’Shaughnessy by a Ph.D. studentship from the Psoriasis Association (Grant No.: ST3 15).
Keywords:SiRNA, cell migration, connexin hemichannel, connexin mimetic peptide, wound healing.
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Johnstone, Dr Scott and Greenhough, Mr Sebastian
Authors: Faniku, C., O'Shaughnessy, E., Lorraine, C., Johnstone, S. R., Graham, A., Greenhough, S., and Martin, P. E.M.
College/School:College of Medical Veterinary and Life Sciences > School of Cardiovascular & Metabolic Health
Journal Name:International Journal of Molecular Sciences
Publisher:MDPI
ISSN:1661-6596
ISSN (Online):1422-0067
Published Online:18 February 2018
Copyright Holders:Copyright © 2018 The Authors
First Published:First published in International Journal of Molecular Sciences 19(2): 604
Publisher Policy:Reproduced under a Creative Commons License

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