Par3 controls actin filament architecture and morphology in epithelial cells of zebrafish skin. (a) Immunofluorescence staining of AmotL2a (green) and F-actin (red) in skin of control (first panel) and par3 MO (second panel) treated zebrafish embryos at 34hpf. In control embryos, AmotL2a localizes to cell-cell junctions, as well as actin positive apical microridges. In the absence of Par3 those actin structures are lost and in addition AmotL2a expression at cell-cell junctions is reduced. The actin filament phenotype could be rescued by co-injecting the morpholino with a human PAR3 mRNA (third panel). (b) As a comparison pictures of skin from amotL2a/b morphants AmotL2a (green) and F-actin (red) show similar to what was observed in the par3 morphants, the embryos deficient of AmotL2 showed loss of cytoplasmic actin filaments. Scale bar 10 um. (c) Quantification of fluorescence intensity of AmotL2 at cell junctions in control and par3 MO injected embryos. N (ctrl) = 100 cells, N (par3 MO) = 100 cells. P values ctrl vs par3 MO < 0.0001 as calculated by unpaired t test. (d) Quantification of cell area in control and par3 MO injected embryos. N (ctrl) = 100 cells, N (par3 MO) = 100 cells. P values ctrl vs par3 MO < 0.0001 as calculated by unpaired t test. (e) Quantification of cell geometry in control, par3 morphants and amotl2 as well as embryos co-injected with par3 MO and human PAR3 mRNA. N (ctrl) = 200 cells, N (par3 MO) = 200, N(amotl2 MO) = 200 cells cells, N(par3 MO + hPAR3 mRNA) = 200 cells. p value ctrl vs. par3 MO < 0.0001. p-value par3 MO vs. par3 MO + hPAR3 mRNA < 0.00001, ctrl vs par3 MO + hPAR3 mRNA = 1. Each experiment was repeated at least three times.
|
