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The PCP genes Celsr1 and Vangl2 are required for normal lung branching morphogenesis.

Yates LL, Schnatwinkel C, Murdoch JN, Bogani D, Formstone CJ, Townsend S, Greenfield A, Niswander LA, Dean CH - Hum. Mol. Genet. (2010)

Bottom Line: Defects in lung architecture are also associated with adult lung disease, particularly in cases of idiopathic lung fibrosis.We observe a recapitulation of these branching defects following inhibition of Rho kinase, an important downstream effector of the PCP signalling pathway.Moreover, epithelial integrity is disrupted, cytoskeletal remodelling perturbed and mutant endoderm does not branch normally in response to the chemoattractant FGF10.

View Article: PubMed Central - PubMed

Affiliation: Medical Research Council, Harwell, Oxfordshire OX11 0RD, UK.

ABSTRACT
The lungs are generated by branching morphogenesis as a result of reciprocal signalling interactions between the epithelium and mesenchyme during development. Mutations that disrupt formation of either the correct number or shape of epithelial branches affect lung function. This, in turn, can lead to congenital abnormalities such as cystadenomatoid malformations, pulmonary hypertension or lung hypoplasia. Defects in lung architecture are also associated with adult lung disease, particularly in cases of idiopathic lung fibrosis. Identifying the signalling pathways which drive epithelial tube formation will likely shed light on both congenital and adult lung disease. Here we show that mutations in the planar cell polarity (PCP) genes Celsr1 and Vangl2 lead to disrupted lung development and defects in lung architecture. Lungs from Celsr1(Crsh) and Vangl2(Lp) mouse mutants are small and misshapen with fewer branches, and by late gestation exhibit thickened interstitial mesenchyme and defective saccular formation. We observe a recapitulation of these branching defects following inhibition of Rho kinase, an important downstream effector of the PCP signalling pathway. Moreover, epithelial integrity is disrupted, cytoskeletal remodelling perturbed and mutant endoderm does not branch normally in response to the chemoattractant FGF10. We further show that Celsr1 and Vangl2 proteins are present in restricted spatial domains within lung epithelium. Our data show that the PCP genes Celsr1 and Vangl2 are required for foetal lung development thereby revealing a novel signalling pathway critical for this process that will enhance our understanding of congenital and adult lung diseases and may in future lead to novel therapeutic strategies.

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Celsr1 and Vangl2 are expressed in all modes of branching. Confocal images of wild-type E14.5 (A, B, G, H) and Celsr1Crsh (C, D) or E12.5 wild-type (E, F) whole lungs immunostained with anti-cytokeratin (A, C, D, G), anti-Celsr1 (B, E, F) or anti-Vangl2 (H) antibodies. Pictures are slices showing a single layer from a z-stack. In wild-type lungs Celsr1 and Vangl2 are expressed in all three branching modes as described by Metzger et al. (34); examples of planar branching (yellow lines in A, C, D, G); domain branching (red lines in A, D, G) and orthogonal branching (white lines in A, C, G) are highlighted. All three modes are present in Celsr1Crsh and Vangl2Lp lungs (C, D and data not shown) indicating the defects in Celsr1 and Vangl2 are not restricted to a specific mode of branching. Scale bars: (A, B) 125 µM plus ×1.2 zoom, (C) 125 µM plus ×1.2 zoom, (D) 125 µM plus ×1.7 zoom. (E, F) 125 µM plus ×6.3 zoom, (G, H) 125 µM plus ×1.7 zoom.
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DDQ104F4: Celsr1 and Vangl2 are expressed in all modes of branching. Confocal images of wild-type E14.5 (A, B, G, H) and Celsr1Crsh (C, D) or E12.5 wild-type (E, F) whole lungs immunostained with anti-cytokeratin (A, C, D, G), anti-Celsr1 (B, E, F) or anti-Vangl2 (H) antibodies. Pictures are slices showing a single layer from a z-stack. In wild-type lungs Celsr1 and Vangl2 are expressed in all three branching modes as described by Metzger et al. (34); examples of planar branching (yellow lines in A, C, D, G); domain branching (red lines in A, D, G) and orthogonal branching (white lines in A, C, G) are highlighted. All three modes are present in Celsr1Crsh and Vangl2Lp lungs (C, D and data not shown) indicating the defects in Celsr1 and Vangl2 are not restricted to a specific mode of branching. Scale bars: (A, B) 125 µM plus ×1.2 zoom, (C) 125 µM plus ×1.2 zoom, (D) 125 µM plus ×1.7 zoom. (E, F) 125 µM plus ×6.3 zoom, (G, H) 125 µM plus ×1.7 zoom.

Mentions: Recent studies identified three modes of branching in mouse lung: domain branching, planar bifurcation and orthogonal bifurcation (34). These three modes of branching are used repeatedly to form a stereotypical pattern of branches in the lung. Domain branching is particularly used during the early stages of branching and involves new bud formation at regular intervals both along the length and around the circumference of an existing branch creating a ‘bottle-brush’ type of structure. Planar and orthogonal bifurcation both involve a single bud tip separating into two distinct tips that are either within the same plane (planar) or at right angles (orthogonal) to the original end bud (see Metzger et al. 34 for further details). To determine whether mutations in the PCP pathway affected all branching modes or whether PCP signalling is part of the molecular mechanisms controlling formation of branches by one particular mode, we immunostained wild-type E12.5 and E14.5 whole lungs with antibodies to Celsr1 or Vangl2 and counterstained with pan-cytokeratin to highlight epithelium (Fig. 4, yellow, red, white lines denote planar, domain and orthogonal branching, respectively). Laser scanning confocal examination of wild-type lungs revealed Celsr1 and Vangl2 expression in all three branch modes (Fig. 4B, E, F and H). In addition, despite the defects already described and in particular the decreased branch number, examples of all three modes of branching were visible in Celsr1Crsh and Vangl2Lp homozygous lungs (Fig. 4C, D and data not shown) providing further evidence that Celsr1 and Vangl2 are part of a general mechanism governing bud/branch formation.


The PCP genes Celsr1 and Vangl2 are required for normal lung branching morphogenesis.

Yates LL, Schnatwinkel C, Murdoch JN, Bogani D, Formstone CJ, Townsend S, Greenfield A, Niswander LA, Dean CH - Hum. Mol. Genet. (2010)

Celsr1 and Vangl2 are expressed in all modes of branching. Confocal images of wild-type E14.5 (A, B, G, H) and Celsr1Crsh (C, D) or E12.5 wild-type (E, F) whole lungs immunostained with anti-cytokeratin (A, C, D, G), anti-Celsr1 (B, E, F) or anti-Vangl2 (H) antibodies. Pictures are slices showing a single layer from a z-stack. In wild-type lungs Celsr1 and Vangl2 are expressed in all three branching modes as described by Metzger et al. (34); examples of planar branching (yellow lines in A, C, D, G); domain branching (red lines in A, D, G) and orthogonal branching (white lines in A, C, G) are highlighted. All three modes are present in Celsr1Crsh and Vangl2Lp lungs (C, D and data not shown) indicating the defects in Celsr1 and Vangl2 are not restricted to a specific mode of branching. Scale bars: (A, B) 125 µM plus ×1.2 zoom, (C) 125 µM plus ×1.2 zoom, (D) 125 µM plus ×1.7 zoom. (E, F) 125 µM plus ×6.3 zoom, (G, H) 125 µM plus ×1.7 zoom.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2865378&req=5

DDQ104F4: Celsr1 and Vangl2 are expressed in all modes of branching. Confocal images of wild-type E14.5 (A, B, G, H) and Celsr1Crsh (C, D) or E12.5 wild-type (E, F) whole lungs immunostained with anti-cytokeratin (A, C, D, G), anti-Celsr1 (B, E, F) or anti-Vangl2 (H) antibodies. Pictures are slices showing a single layer from a z-stack. In wild-type lungs Celsr1 and Vangl2 are expressed in all three branching modes as described by Metzger et al. (34); examples of planar branching (yellow lines in A, C, D, G); domain branching (red lines in A, D, G) and orthogonal branching (white lines in A, C, G) are highlighted. All three modes are present in Celsr1Crsh and Vangl2Lp lungs (C, D and data not shown) indicating the defects in Celsr1 and Vangl2 are not restricted to a specific mode of branching. Scale bars: (A, B) 125 µM plus ×1.2 zoom, (C) 125 µM plus ×1.2 zoom, (D) 125 µM plus ×1.7 zoom. (E, F) 125 µM plus ×6.3 zoom, (G, H) 125 µM plus ×1.7 zoom.
Mentions: Recent studies identified three modes of branching in mouse lung: domain branching, planar bifurcation and orthogonal bifurcation (34). These three modes of branching are used repeatedly to form a stereotypical pattern of branches in the lung. Domain branching is particularly used during the early stages of branching and involves new bud formation at regular intervals both along the length and around the circumference of an existing branch creating a ‘bottle-brush’ type of structure. Planar and orthogonal bifurcation both involve a single bud tip separating into two distinct tips that are either within the same plane (planar) or at right angles (orthogonal) to the original end bud (see Metzger et al. 34 for further details). To determine whether mutations in the PCP pathway affected all branching modes or whether PCP signalling is part of the molecular mechanisms controlling formation of branches by one particular mode, we immunostained wild-type E12.5 and E14.5 whole lungs with antibodies to Celsr1 or Vangl2 and counterstained with pan-cytokeratin to highlight epithelium (Fig. 4, yellow, red, white lines denote planar, domain and orthogonal branching, respectively). Laser scanning confocal examination of wild-type lungs revealed Celsr1 and Vangl2 expression in all three branch modes (Fig. 4B, E, F and H). In addition, despite the defects already described and in particular the decreased branch number, examples of all three modes of branching were visible in Celsr1Crsh and Vangl2Lp homozygous lungs (Fig. 4C, D and data not shown) providing further evidence that Celsr1 and Vangl2 are part of a general mechanism governing bud/branch formation.

Bottom Line: Defects in lung architecture are also associated with adult lung disease, particularly in cases of idiopathic lung fibrosis.We observe a recapitulation of these branching defects following inhibition of Rho kinase, an important downstream effector of the PCP signalling pathway.Moreover, epithelial integrity is disrupted, cytoskeletal remodelling perturbed and mutant endoderm does not branch normally in response to the chemoattractant FGF10.

View Article: PubMed Central - PubMed

Affiliation: Medical Research Council, Harwell, Oxfordshire OX11 0RD, UK.

ABSTRACT
The lungs are generated by branching morphogenesis as a result of reciprocal signalling interactions between the epithelium and mesenchyme during development. Mutations that disrupt formation of either the correct number or shape of epithelial branches affect lung function. This, in turn, can lead to congenital abnormalities such as cystadenomatoid malformations, pulmonary hypertension or lung hypoplasia. Defects in lung architecture are also associated with adult lung disease, particularly in cases of idiopathic lung fibrosis. Identifying the signalling pathways which drive epithelial tube formation will likely shed light on both congenital and adult lung disease. Here we show that mutations in the planar cell polarity (PCP) genes Celsr1 and Vangl2 lead to disrupted lung development and defects in lung architecture. Lungs from Celsr1(Crsh) and Vangl2(Lp) mouse mutants are small and misshapen with fewer branches, and by late gestation exhibit thickened interstitial mesenchyme and defective saccular formation. We observe a recapitulation of these branching defects following inhibition of Rho kinase, an important downstream effector of the PCP signalling pathway. Moreover, epithelial integrity is disrupted, cytoskeletal remodelling perturbed and mutant endoderm does not branch normally in response to the chemoattractant FGF10. We further show that Celsr1 and Vangl2 proteins are present in restricted spatial domains within lung epithelium. Our data show that the PCP genes Celsr1 and Vangl2 are required for foetal lung development thereby revealing a novel signalling pathway critical for this process that will enhance our understanding of congenital and adult lung diseases and may in future lead to novel therapeutic strategies.

Show MeSH
Related in: MedlinePlus