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Fig. 6. Vangl2 and aPKC are required for specific oocyte membrane and mRNA asymmetries. (A,A′) Low and high magnification images of hemisected whole-mount, progesterone-matured control (A) and Vangl2 depleted (VG–; A′) Xenopus oocytes stained for endogenous aPKC. (B,C) Localization of overexpressed GFP-LGL protein in anti-GFP immunostained histological sections of a stage 6 control oocyte (B) and progesterone-matured control and aPKC-depleted (PKC–) oocytes injected with GFP-LGL mRNA (C). Insets show bright-field images. Chevrons indicate the boundary between the animal and the vegetal hemispheres. (D) Comparison of the staining pattern of GFP-LGL in whole-mount anti-GFP immunostained control and Vangl2-depleted (VG–) progesterone-matured oocytes. Insets show the brightfield appearance of each oocyte and chevrons indicate the junction between animal and vegetal hemispheres. (E) Control for experiment shown in D showing a progesterone-matured oocyte that did not receive GFP-LGL mRNA injection. (F) GFP-LGL protein localization in histological sections of control, aPKC-depleted (aPKC–) and Vangl2-depleted (VG–) animal cells of embryos at the blastula stage, injected with GFP-LGL mRNA at the 2-cell stage. Chevrons mark the apical membrane of superficial cells in the animal cap. (G) In situ hybridization of sibling control, Vangl2-depleted (VG–) and aPKC-depleted (aPKC–) stage 6 oocytes probed for Wnt11, VegT and Xpat mRNAs. (H) Relative expression levels of Wnt11, VegT and Xpat mRNAs in control (uninj oocyte), Vangl2-depleted (VG–) and aPKC-depleted (aPKC–) oocytes determined by real-time RT-PCR (mean±s.d.). |
