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To express EPS-8 isoforms in embryonic dorsal and ventral epidermal cells and in pharyngeal marginal cells we used the vab-19 promoter.
For all the marginal cells we tested (n = 8), the dye stayed in the injected cell for more than 30 minutes (Fig. 3D).
To quantitatively assess gene transfection efficiency in the marginal cells, we calculated the percentage of transduced cells by counting the number of Kcnq1-positive cells (Fig 3) and then divided the number by the total number of marginal cells in the field of view.
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Zebrafish Wnt11 expresses at the epiblast cells of the germ ring, which correspond to the position of marginal deep cells we observed in this study.
Since one of the aims of this study was to re-establish the missing Kcnq1 expression (Fig 1C) in as many marginal cells as possible, we first compared the viral inoculation efficiency achieved by injections into the SM and scala tympani (ST).
To assess whether the expansion of the scala media in Atp6v0a4 mutants affects the integrity and function of marginal cells of the stria vascularis, we examined the expression of Kcnq1 in Atp6v0a4 −/− and controls at different time points.
We found that marginal cells in tissue explants not only rapidly transformed into stable-bleb cells, as expected for LPA-treated progenitors, but also frequently extruded from the explant surface.
As a negative control, we injected PI into the marginal cells since these cells are not coupled by GJs [13], [14].
To determine whether EPS-8 localizes to a specific part of cell-matrix attachments we examined attachment structures of pharyngeal marginal cells.
We measured the length of the marginal cells and of terminal oocytes of these bees on day 7.
We also found that, compared to the marginal cells in WT mice (Fig 3E), the sizes of the marginal cells in untreated Kcnq1−/− mice vary greatly.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com