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. 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 n = 37 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 10 μm) of selected time-points of the wounding assay shown in a, c. Targeted area (95 μm2) 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 (22 μm) displayed on the 0 s, UtrCH micrograph. The cell shown in a, c, and d is representative of n = 37 wounding assays, all of which were used to prepare b. Scale bars = 10 μm
	Fig. 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 n = 12 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 10 μm) of selected time-points of the wounding assay shown in a, c. Targeted area (95 μm2) 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 (22 μm) displayed on the 0 s, MRLC micrograph. The cell shown in a, c, and d is representative of n = 12 wounding assays, all of which were used to prepare b. Scale bars = 10 μm
	Fig. 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 γGBD signals following laser-mediated ablation of the PM of HeLa cells. b Normalized γGBD fluorescent signal intensity (γGBD FIR) at the wound site relative to control regions. Mean and SEM shown of n = 12 cells. c Kymographs of the UtrCH and γGBD 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 (95 μm2) 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 (22 μm) displayed on the 0 s, γGBD micrograph. The cell shown in a, c is representative of n = 12 wounding assays, all of which were used to prepare b. Scale bars = 10 μm

References [+] :

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