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Wnt ligands from the embryonic surface ectoderm regulate 'bimetallic strip' optic cup morphogenesis in mouse.

Carpenter AC, Smith AN, Wagner H, Cohen-Tayar Y, Rao S, Wallace V, Ashery-Padan R, Lang RA - Development (2015)

Bottom Line: Consistent with this, microarray and cell fate marker analysis identified a series of expression changes in genes known to be regulated by RA or by the Wnt/β-catenin pathway.Using pathway reporters, we showed that Wnt ligands from the surface ectoderm directly or indirectly elicit a Wnt/β-catenin response in retinal pigment epithelium (RPE) progenitors near the optic cup rim.These data thus establish a novel hypothesis to explain how differential cell numbers in a bilayered epithelium can lead to shape change.

View Article: PubMed Central - PubMed

Affiliation: Visual Systems Group, Abrahamson Pediatric Eye Institute, Division of Pediatric Ophthalmology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.

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Related in: MedlinePlus

Flow sorting and Wnt expression in ocular ectoderm and mesenchyme. (A,C) GFP labeling in transverse cryosections of the eye region from Le-cre; Z/EG (A) and Wnt1-cre; Z/EG (C) mouse embryos at E10.5. Hoechst 33258 nuclear staining is in blue. (B,D) Flow sorting of GFP+ cells (green) from Le-cre; Z/EG (B) and Wnt1-cre; Z/EG (D) eye tissue at E10.5. (E) End-point PCR for labeled Wnt transcripts from flow-sorted GFP+ cells of the indicated genotype. Red dots indicate bands of the correct size for the particular Wnt transcript; red crosses indicate bands of incorrect size.
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DEV120022F1: Flow sorting and Wnt expression in ocular ectoderm and mesenchyme. (A,C) GFP labeling in transverse cryosections of the eye region from Le-cre; Z/EG (A) and Wnt1-cre; Z/EG (C) mouse embryos at E10.5. Hoechst 33258 nuclear staining is in blue. (B,D) Flow sorting of GFP+ cells (green) from Le-cre; Z/EG (B) and Wnt1-cre; Z/EG (D) eye tissue at E10.5. (E) End-point PCR for labeled Wnt transcripts from flow-sorted GFP+ cells of the indicated genotype. Red dots indicate bands of the correct size for the particular Wnt transcript; red crosses indicate bands of incorrect size.

Mentions: Wnt responses are implicated in early eye development (Smith et al., 2005; Fuhrmann, 2008; Swindell et al., 2008; Zhou et al., 2008). To define the ligand genes expressed in eye tissues that were Cre targetable, we flow sorted GFP+ cells from Z/EG mice (Novak et al., 2000) converted by Le-cre (Ashery-Padan et al., 2000) and Wnt1-cre (Brault et al., 2001); these identify, respectively, ocular ectoderm and neural crest-derived POM cells according to antibody labeling for GFP in cryosections (Fig. 1A,C). Flow sorting of GFP+ cells from dissected eye regions from E10.5 showed well separated populations (Fig. 1B,D). End-point RT-PCR on mRNA isolated from these GFP+ populations showed that many Wnt ligands were expressed. Ectoderm expressing Le-cre expressed the majority of the Wnt ligand family, including Wnt1, 2, 3, 3a, 4, 5a, 6, 9b, 10a, 10b and 11 (Fig 1E). Sorted neural crest-derived POM cells expressed mRNA for Wnt2b, 3, 3a, 5a, 7b, 9a, 9b, 10 and 11 (Fig 1E).Fig. 1.


Wnt ligands from the embryonic surface ectoderm regulate 'bimetallic strip' optic cup morphogenesis in mouse.

Carpenter AC, Smith AN, Wagner H, Cohen-Tayar Y, Rao S, Wallace V, Ashery-Padan R, Lang RA - Development (2015)

Flow sorting and Wnt expression in ocular ectoderm and mesenchyme. (A,C) GFP labeling in transverse cryosections of the eye region from Le-cre; Z/EG (A) and Wnt1-cre; Z/EG (C) mouse embryos at E10.5. Hoechst 33258 nuclear staining is in blue. (B,D) Flow sorting of GFP+ cells (green) from Le-cre; Z/EG (B) and Wnt1-cre; Z/EG (D) eye tissue at E10.5. (E) End-point PCR for labeled Wnt transcripts from flow-sorted GFP+ cells of the indicated genotype. Red dots indicate bands of the correct size for the particular Wnt transcript; red crosses indicate bands of incorrect size.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4352985&req=5

DEV120022F1: Flow sorting and Wnt expression in ocular ectoderm and mesenchyme. (A,C) GFP labeling in transverse cryosections of the eye region from Le-cre; Z/EG (A) and Wnt1-cre; Z/EG (C) mouse embryos at E10.5. Hoechst 33258 nuclear staining is in blue. (B,D) Flow sorting of GFP+ cells (green) from Le-cre; Z/EG (B) and Wnt1-cre; Z/EG (D) eye tissue at E10.5. (E) End-point PCR for labeled Wnt transcripts from flow-sorted GFP+ cells of the indicated genotype. Red dots indicate bands of the correct size for the particular Wnt transcript; red crosses indicate bands of incorrect size.
Mentions: Wnt responses are implicated in early eye development (Smith et al., 2005; Fuhrmann, 2008; Swindell et al., 2008; Zhou et al., 2008). To define the ligand genes expressed in eye tissues that were Cre targetable, we flow sorted GFP+ cells from Z/EG mice (Novak et al., 2000) converted by Le-cre (Ashery-Padan et al., 2000) and Wnt1-cre (Brault et al., 2001); these identify, respectively, ocular ectoderm and neural crest-derived POM cells according to antibody labeling for GFP in cryosections (Fig. 1A,C). Flow sorting of GFP+ cells from dissected eye regions from E10.5 showed well separated populations (Fig. 1B,D). End-point RT-PCR on mRNA isolated from these GFP+ populations showed that many Wnt ligands were expressed. Ectoderm expressing Le-cre expressed the majority of the Wnt ligand family, including Wnt1, 2, 3, 3a, 4, 5a, 6, 9b, 10a, 10b and 11 (Fig 1E). Sorted neural crest-derived POM cells expressed mRNA for Wnt2b, 3, 3a, 5a, 7b, 9a, 9b, 10 and 11 (Fig 1E).Fig. 1.

Bottom Line: Consistent with this, microarray and cell fate marker analysis identified a series of expression changes in genes known to be regulated by RA or by the Wnt/β-catenin pathway.Using pathway reporters, we showed that Wnt ligands from the surface ectoderm directly or indirectly elicit a Wnt/β-catenin response in retinal pigment epithelium (RPE) progenitors near the optic cup rim.These data thus establish a novel hypothesis to explain how differential cell numbers in a bilayered epithelium can lead to shape change.

View Article: PubMed Central - PubMed

Affiliation: Visual Systems Group, Abrahamson Pediatric Eye Institute, Division of Pediatric Ophthalmology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.

Show MeSH
Related in: MedlinePlus