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Mutant p63 causes defective expansion of ectodermal progenitor cells and impaired FGF signalling in AEC syndrome.

Ferone G, Thomason HA, Antonini D, De Rosa L, Hu B, Gemei M, Zhou H, Ambrosio R, Rice DP, Acampora D, van Bokhoven H, Del Vecchio L, Koster MI, Tadini G, Spencer-Dene B, Dixon M, Dixon J, Missero C - EMBO Mol Med (2012)

Bottom Line: The AEC mutation exerts a selective dominant-negative function on wild-type p63 by affecting progenitor cell expansion during ectodermal development leading to a defective epidermal stem cell compartment.These phenotypes are associated with impairment of fibroblast growth factor (FGF) signalling resulting from reduced expression of Fgfr2 and Fgfr3, direct p63 target genes.Restoring Fgfr2b expression in p63(+/L514F) epithelial cells by treatment with FGF7 reactivates downstream mitogen-activated protein kinase signalling and cell proliferation.

View Article: PubMed Central - PubMed

Affiliation: Fondazione IRCCS SDN, Napoli, Italy.

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Palatal shelf hypoplasia and reduced epithelial cell proliferation in p63+/L514F embryosH&E staining of coronal sections of p63+/L514F mid palate at E13.5 reveals grossly similar morphology of palatal shelves compared to controls. t, tongue; p, palatal shelf; tb, molar tooth bud; m, mandible. Scale bar: 200 µm.H&E and Alcian Blue staining of coronal sections of p63+/L514F mid palate at E14.5 reveals a failure of the palatal shelves to meet in the horizontal plane. nc, nasal cavity. Scale bar: 200 µm.A closer view of H&E staining of E13.5 coronal sections reveals that mutant palatal shelves are hypoplastic compared to controls. Scale bar: 50 µm.Quantification of the anterior, mid and posterior palatal shelf area. Error bars represent standard deviation (SD). Anterior palate *p-value = 0.00032; n = 9. Mid palate *p-value = 0.00079; n = 9. Posterior palate p-value = 0.0097; n = 9.Immunofluorescence with anti-BrdU antibodies at E13 (left panel) reveals reduced proliferation of mutant palatal shelves.The average percentage of BrdU incorporation is reduced in mutant palatal epithelium at E13 and E13.5. Epithelium at E13 p-value = 0.009; n = 9. Mesenchyme at E13 p-value = 0.032; n = 9. Epithelium at E13.5 p-value = 0.018; n = 9. Data are represented as mean ± SD. Scale bar: 50 µm.Double immunofluorescence staining for the indicated markers reveals hypoplasia and disorganization of the palatal shelf epithelium at E13.5. Scale bar: 20 µm.Palatal explant culture assay reveals that mutant palatal shelves fuse appropriately. Palatal shelves freshly isolated at E13.5 were placed in contact and cultured for 48 h. ne, nasal epithelium; oe, oral epithelium.
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fig02: Palatal shelf hypoplasia and reduced epithelial cell proliferation in p63+/L514F embryosH&E staining of coronal sections of p63+/L514F mid palate at E13.5 reveals grossly similar morphology of palatal shelves compared to controls. t, tongue; p, palatal shelf; tb, molar tooth bud; m, mandible. Scale bar: 200 µm.H&E and Alcian Blue staining of coronal sections of p63+/L514F mid palate at E14.5 reveals a failure of the palatal shelves to meet in the horizontal plane. nc, nasal cavity. Scale bar: 200 µm.A closer view of H&E staining of E13.5 coronal sections reveals that mutant palatal shelves are hypoplastic compared to controls. Scale bar: 50 µm.Quantification of the anterior, mid and posterior palatal shelf area. Error bars represent standard deviation (SD). Anterior palate *p-value = 0.00032; n = 9. Mid palate *p-value = 0.00079; n = 9. Posterior palate p-value = 0.0097; n = 9.Immunofluorescence with anti-BrdU antibodies at E13 (left panel) reveals reduced proliferation of mutant palatal shelves.The average percentage of BrdU incorporation is reduced in mutant palatal epithelium at E13 and E13.5. Epithelium at E13 p-value = 0.009; n = 9. Mesenchyme at E13 p-value = 0.032; n = 9. Epithelium at E13.5 p-value = 0.018; n = 9. Data are represented as mean ± SD. Scale bar: 50 µm.Double immunofluorescence staining for the indicated markers reveals hypoplasia and disorganization of the palatal shelf epithelium at E13.5. Scale bar: 20 µm.Palatal explant culture assay reveals that mutant palatal shelves fuse appropriately. Palatal shelves freshly isolated at E13.5 were placed in contact and cultured for 48 h. ne, nasal epithelium; oe, oral epithelium.

Mentions: To identify the cause of cleft palate, we analyzed the p63+/L514F mouse oral cavity during development. At embryonic day (E)13.5, although reproducibly smaller, the mutant palatal shelves pointed downwards on either side of the tongue similar to their wild-type littermates (Fig 2A, C and D). Although the mutant palatal shelves elevated appropriately at E14.5, they were smaller, widely spaced and failed to meet in the midline (Fig 2B).


Mutant p63 causes defective expansion of ectodermal progenitor cells and impaired FGF signalling in AEC syndrome.

Ferone G, Thomason HA, Antonini D, De Rosa L, Hu B, Gemei M, Zhou H, Ambrosio R, Rice DP, Acampora D, van Bokhoven H, Del Vecchio L, Koster MI, Tadini G, Spencer-Dene B, Dixon M, Dixon J, Missero C - EMBO Mol Med (2012)

Palatal shelf hypoplasia and reduced epithelial cell proliferation in p63+/L514F embryosH&E staining of coronal sections of p63+/L514F mid palate at E13.5 reveals grossly similar morphology of palatal shelves compared to controls. t, tongue; p, palatal shelf; tb, molar tooth bud; m, mandible. Scale bar: 200 µm.H&E and Alcian Blue staining of coronal sections of p63+/L514F mid palate at E14.5 reveals a failure of the palatal shelves to meet in the horizontal plane. nc, nasal cavity. Scale bar: 200 µm.A closer view of H&E staining of E13.5 coronal sections reveals that mutant palatal shelves are hypoplastic compared to controls. Scale bar: 50 µm.Quantification of the anterior, mid and posterior palatal shelf area. Error bars represent standard deviation (SD). Anterior palate *p-value = 0.00032; n = 9. Mid palate *p-value = 0.00079; n = 9. Posterior palate p-value = 0.0097; n = 9.Immunofluorescence with anti-BrdU antibodies at E13 (left panel) reveals reduced proliferation of mutant palatal shelves.The average percentage of BrdU incorporation is reduced in mutant palatal epithelium at E13 and E13.5. Epithelium at E13 p-value = 0.009; n = 9. Mesenchyme at E13 p-value = 0.032; n = 9. Epithelium at E13.5 p-value = 0.018; n = 9. Data are represented as mean ± SD. Scale bar: 50 µm.Double immunofluorescence staining for the indicated markers reveals hypoplasia and disorganization of the palatal shelf epithelium at E13.5. Scale bar: 20 µm.Palatal explant culture assay reveals that mutant palatal shelves fuse appropriately. Palatal shelves freshly isolated at E13.5 were placed in contact and cultured for 48 h. ne, nasal epithelium; oe, oral epithelium.
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Related In: Results  -  Collection

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fig02: Palatal shelf hypoplasia and reduced epithelial cell proliferation in p63+/L514F embryosH&E staining of coronal sections of p63+/L514F mid palate at E13.5 reveals grossly similar morphology of palatal shelves compared to controls. t, tongue; p, palatal shelf; tb, molar tooth bud; m, mandible. Scale bar: 200 µm.H&E and Alcian Blue staining of coronal sections of p63+/L514F mid palate at E14.5 reveals a failure of the palatal shelves to meet in the horizontal plane. nc, nasal cavity. Scale bar: 200 µm.A closer view of H&E staining of E13.5 coronal sections reveals that mutant palatal shelves are hypoplastic compared to controls. Scale bar: 50 µm.Quantification of the anterior, mid and posterior palatal shelf area. Error bars represent standard deviation (SD). Anterior palate *p-value = 0.00032; n = 9. Mid palate *p-value = 0.00079; n = 9. Posterior palate p-value = 0.0097; n = 9.Immunofluorescence with anti-BrdU antibodies at E13 (left panel) reveals reduced proliferation of mutant palatal shelves.The average percentage of BrdU incorporation is reduced in mutant palatal epithelium at E13 and E13.5. Epithelium at E13 p-value = 0.009; n = 9. Mesenchyme at E13 p-value = 0.032; n = 9. Epithelium at E13.5 p-value = 0.018; n = 9. Data are represented as mean ± SD. Scale bar: 50 µm.Double immunofluorescence staining for the indicated markers reveals hypoplasia and disorganization of the palatal shelf epithelium at E13.5. Scale bar: 20 µm.Palatal explant culture assay reveals that mutant palatal shelves fuse appropriately. Palatal shelves freshly isolated at E13.5 were placed in contact and cultured for 48 h. ne, nasal epithelium; oe, oral epithelium.
Mentions: To identify the cause of cleft palate, we analyzed the p63+/L514F mouse oral cavity during development. At embryonic day (E)13.5, although reproducibly smaller, the mutant palatal shelves pointed downwards on either side of the tongue similar to their wild-type littermates (Fig 2A, C and D). Although the mutant palatal shelves elevated appropriately at E14.5, they were smaller, widely spaced and failed to meet in the midline (Fig 2B).

Bottom Line: The AEC mutation exerts a selective dominant-negative function on wild-type p63 by affecting progenitor cell expansion during ectodermal development leading to a defective epidermal stem cell compartment.These phenotypes are associated with impairment of fibroblast growth factor (FGF) signalling resulting from reduced expression of Fgfr2 and Fgfr3, direct p63 target genes.Restoring Fgfr2b expression in p63(+/L514F) epithelial cells by treatment with FGF7 reactivates downstream mitogen-activated protein kinase signalling and cell proliferation.

View Article: PubMed Central - PubMed

Affiliation: Fondazione IRCCS SDN, Napoli, Italy.

Show MeSH
Related in: MedlinePlus