Dominant effect of gap junction communication in wound-induced calcium-wave, NFAT activation and wound closure in keratinocytes
| Autor: | Malcolm Begg, Colin A.B. Jahoda, Timothy R. Cheek, Laura Hudson, Nick J. Reynolds, Blythe Wright |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Keratinocytes Physiology Clinical Biochemistry chemistry.chemical_element Calcium 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Adenosine Triphosphate Cell Movement Calcium flux Animals Humans Channel blocker Calcium Signaling Cells Cultured Hexokinase Wound Healing NFATC Transcription Factors Chemistry Gap Junctions NFAT Cell migration Cell Biology Store-operated calcium entry 030104 developmental biology 030220 oncology & carcinogenesis Biophysics Wound healing |
| Zdroj: | Journal of Cellular Physiology, 2021, Vol.236(12), pp.8171-8183 [Peer Reviewed Journal] |
| ISSN: | 1097-4652 |
| Popis: | Wounding induces a calcium wave and disrupts the calcium gradient across the epidermis but mechanisms mediating calcium and downstream signalling, and longer-term wound healing responses are incompletely understood. As expected, live-cell confocal imaging of Fluo-4-loaded normal human keratinocytes showed an immediate increase in [Ca2+ ]i at the wound edge that spread as a calcium wave (8.3 µm/s) away from the wound edge with gradually diminishing rate of rise and amplitude. The amplitude and area under the curve of [Ca2+ ]i flux was increased in high (1.2 mM) [Ca2+ ]o media. 18α-glycyrrhetinic acid (18αGA), a gap-junction inhibitor or hexokinase, an ATP scavenger, blocked the wound-induced calcium wave, dependent in part on [Ca2+ ]o . Wounding in a high [Ca2+ ]o increased nuclear factor of activated T-cells (NFAT) but not NFkB activation, assessed by dual-luciferase receptor assays compared to unwounded cells. Treatment with 18αGA or the store-operated channel blocker GSK-7975A inhibited wound-induced NFAT activation, whereas treatment with hexokinase did not. Real-time cell migration analysis, measuring wound closure rates over 24 h, revealed that 18αGA essentially blocked wound closure whereas hexokinase and GSK-7975A showed relatively minimal effects. Together these data indicate that while both gap-junction communication and ATP release from damaged cells are important in regulating the wound-induced calcium wave, long-term transcriptional and functional responses are dominantly regulated by gap-junction communication. |
| Databáze: | OpenAIRE |
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