DeKraker C et al. (2019), Dynamics of actin polymerisation during the mam...

XB-ART-56123

BMC Res Notes 2019 Jul 16;121:420. doi: 10.1186/s13104-019-4441-7.

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Dynamics of actin polymerisation during the mammalian single-cell wound healing response.

DeKraker C , Goldin-Blais L , Boucher E , Mandato CA .

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OBJECTIVE: The contribution of actomyosin contractile rings in the wound healing program of somatic cells as never been directly assessed. This contrast with the events characterising the wound healing response of in wounded Xenopus oocytes, in which formation and contraction of an actomyosin ring provides a platform for cytoskeletal repair and drives the restoration of proper plasma membrane composition at the site of injury. As such, we aimed to characterize, using high-resolution live-cell confocal microscopy, the cytoskeletal repair dynamics of HeLa cells. RESULTS: We confirm here that the F-actin enrichment that characterizes the late repair program of laser-wounded cells is mostly uniform and is not associated with co-enrichment of myosin-II or the formation of concentric zones of RhoA and Cdc42 activity.

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Species referenced: Xenopus
Genes referenced: cdc42 rhoa

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	Fig.Â 1. F-actin is enriched at the wound site following laser-mediated ablation of the PM. a Maximum intensity projections of selected micrographs of the CellMask and UtrCH signals following laser-mediated ablation of the PM of HeLa cells. b Normalized UtrCH fluorescent signal intensity (UtrCH FIR) at the wound site relative to control regions. Mean and SEM shown of nâ=â37 cells. c Kymograph of the UtrCH signal before and after laser-mediated ablation of the PM. Cell and targeted area is the same as in the one shown in a. d Resliced cortical volume (each 10Â Î¼m) of selected time-points of the wounding assay shown in a, c. Targeted area (95Â Î¼m2) is represented by the red circle found on the pre-wounded, PM micrograph. The line scan region used for the creation of the kymograph and resliced volumes correspond to the cyan line (22Â Î¼m) displayed on the 0Â s, UtrCH micrograph. The cell shown in a, c, and d is representative of nâ=â37 wounding assays, all of which were used to prepare b. Scale barsâ=â10Â Î¼m
	Fig.Â 2. Mammalian single-cell repair does not involve myosin enrichment at the wound borders. a Maximum intensity projections of selected micrographs of the CellMask and MRLC signals following laser-mediated ablation of the PM of HeLa cells. b Normalized MLRC fluorescent signal intensity (MRLC FIR) at the wound site relative to control regions. Mean and SEM shown of nâ=â12 cells. c Kymograph of the MRLC signal before and after laser-mediated ablation of the PM. Cell and targeted area is the same as in the one shown in a. d Resliced cortical volume (each 10Â Î¼m) of selected time-points of the wounding assay shown in a, c. Targeted area (95Â Î¼m2) is represented by the red circle found on the pre-wounded, PM micrograph. The line scan region used for the creation of the kymograph and resliced volumes correspond to the cyan line (22Â Î¼m) displayed on the 0Â s, MRLC micrograph. The cell shown in a, c, and d is representative of nâ=â12 wounding assays, all of which were used to prepare b. Scale barsâ=â10Â Î¼m
	Fig.Â 3. RhoA activity is increased the over entire area of the wound-site. a Maximum intensity projections of selected micrographs of the CellMask, UtrCH, and Î³GBD signals following laser-mediated ablation of the PM of HeLa cells. b Normalized Î³GBD fluorescent signal intensity (Î³GBD FIR) at the wound site relative to control regions. Mean and SEM shown of nâ=â12 cells. c Kymographs of the UtrCH and Î³GBD signals before and after laser-mediated ablation of the PM. Cell and targeted area is the same as in the one shown in a. Targeted area (95Â Î¼m2) is represented by the red circle found on the pre-wounded, PM micrograph. The line scan region used for the creation of the kymograph correspond to the cyan line (22Â Î¼m) displayed on the 0Â s, Î³GBD micrograph. The cell shown in a, c is representative of nâ=â12 wounding assays, all of which were used to prepare b. Scale barsâ=â10Â Î¼m

References [+] :

Abreu-Blanco, Cell wound repair in Drosophila occurs through three distinct phases of membrane and cytoskeletal remodeling. 2011, Pubmed