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Shalini Shatadal, Rivka Rachel, Anne Griep; Dlg-1 and Scrib are Modulators of Wnt/PCP in the Mouse Ocular Lens. Invest. Ophthalmol. Vis. Sci. 2012;53(14):5594.
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© ARVO (1962-2015); The Authors (2016-present)
Previously, we reported that Dlg-1 and Scrib, mouse homologs of Drosophila tumor suppressors, dlg and scrib, are required for mouse lens development. Furthermore, we recently showed that these genes interact with Vangl2 (a core member of Wnt/PCP pathway), to modulate cell adhesion and apical-basal polarity in the epithelium. The lens has been shown to exhibit PCP in the outer cortical fiber cells. In this study we ask if Dlg-1 and Scrib, through their interaction with Vangl2, modulate PCP in the lens.
Dlg-1 or Scrib conditional null mice were crossed with MLR10cre mice and the progeny crossed with Vangl2Lp/+ mice. Eyes from postnatal day 30 Dlgf/+, Dlgf/+10, Vangl2Lp/+;Dlgf/+, Vangl2Lp/+;Dlgf/+10, Scribf/+, Scribf/+10, Vangl2Lp/+;Scribf/+ and Vangl2 Lp/+;Scribf/+10 mice were fixed in 4% PFA, cryoprotected in sucrose, embedded transversely in OCT and 10μm sections were prepared. Immunofluorescence was performed on sections near the equator to assess the localization pattern of PCP proteins Frizzled6 (Fz6), Vangl2, Prickle (Pk) and Dishevelled2 (Dvl2) in the outer cortical fibers. Sections were co-immunostained for N-cadherin to visualize the cell membranes.
The fibers of control Dlgf/+ and Scribf/+ lenses were hexagonally shaped and oriented toward the anterior pole. In contrast, Vangl2Lp/+ and Dlgf/+10 fibers were square or rounded and oriented randomly. Fibers in the Scribf/+10 lens were only mildly distorted. Cell structure in the Vangl2Lp/+;Dlgf/+10 lenses was more disrupted than in Vangl2Lp/+ alone whereas cell structure in Vangl2Lp/+Scribf/+10 fibers was more similar to controls. In the fibers of control lenses, Fz6 and Vangl2 accumulated on the apical surface whereas Pk accumulated on the long sides. Dvl2 staining was found on the long sides and on the apical surface. In contrast, Vangl2 was distributed throughout the membrane in the Dlgf/+10 fibers and some staining on the long sides was observed in Scribf/+10 fibers. In Dlgf/+10, Vangl2Lp/+, and Vangl2Lp/+;Dlgf/+10 fibers, Fz6 was found throughout the membranes while Vangl2, Pk and Dvl2 staining was mislocalized and reduced in intensity. Fz6 and Pk were mislocalized in Scribf/+10 fibers while their localization in Vangl2Lp/+;Scribf/+10 fibers was similar to controls. Dvl2 localization in Scribf/+10 was similar to controls but was partially mislocalized or absent in some areas in Vangl2Lp/+;Scribf/+10 fibers.
These data indicate that Dlg-1 and Scrib are modulators of cell shape, organization and localization of PCP proteins in lens fibers. As heterozygosity for Dlg-1 enhances whereas heterozygosity for Scrib suppresses the Vangl2 phenotype, Dlg-1 and Scrib appear to act in opposing ways with respect to Vangl2 in modulating PCP in the lens fibers.
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