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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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Mutant lung endoderm responds to FGF10 but is unable to undergo branching. (A, B) Wild-type lung endoderm branches in response to 400 ng/ml FGF10 (mean bud number = 11, n = 6). (C, D) mutant lungs do not branch in response to FGF10 (mean bud number Crsh = 0.9, n = 6). P-ERK1/2 staining of E14.5 lung, shows no significant difference in percentage of P-ERK1/2 positive cells in the epithelium of mutants compared with wild-type controls (E): wild-type 55.17%, ±3, n = 10; Crsh 51.97%, ±6.3, n = 7, P = 0.6074; Lp 56.45%, ±3.9, n = 9, P = 0.7939. Data from three embryos for each genotype. Western blotting reveals no change in the relative levels of P-ERK1/2 in wild-type and Lp/Lp or Crsh/Crsh littermates (F). Scale bars: (A–D) 5 µM.
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DDQ104F6: Mutant lung endoderm responds to FGF10 but is unable to undergo branching. (A, B) Wild-type lung endoderm branches in response to 400 ng/ml FGF10 (mean bud number = 11, n = 6). (C, D) mutant lungs do not branch in response to FGF10 (mean bud number Crsh = 0.9, n = 6). P-ERK1/2 staining of E14.5 lung, shows no significant difference in percentage of P-ERK1/2 positive cells in the epithelium of mutants compared with wild-type controls (E): wild-type 55.17%, ±3, n = 10; Crsh 51.97%, ±6.3, n = 7, P = 0.6074; Lp 56.45%, ±3.9, n = 9, P = 0.7939. Data from three embryos for each genotype. Western blotting reveals no change in the relative levels of P-ERK1/2 in wild-type and Lp/Lp or Crsh/Crsh littermates (F). Scale bars: (A–D) 5 µM.

Mentions: We wished to determine whether the tissue morphogenesis defects observed in Celsr1 and Vangl2 mutant lungs could reflect a defect in the response of mutant endoderm to a key signal for branching. To test this idea we exposed Celsr1Crsh, Vangl2Lp and wild-type lung epithelium denuded of mesenchyme to FGF10, which normally signals from the mesenchyme to direct lung branching. Both wild-type and mutant lungs responded to the FGF10 stimulus in terms of growth. However, whereas wild-type epithelium formed multiple long and narrow buds (Fig. 6A and B), in most cases, the mutant lung epithelium did not undergo any branching in response to FGF10 (Fig. 6C, D or rarely one or two short stumpy buds formed). Staining for phospho-ERK1/2, which is up-regulated in response to FGF10 in the lung (40), revealed no significant difference in the number of positive cells in wild-type and mutant lung epithelium and this was confirmed by western blot (Fig. 6E and F). Together these results indicate that the FGF signalling pathway, at least that mediated by ERK1/2 activators, is unaffected in mutant lungs. Thus, normal morphogenetic movement within the epithelium is affected upon disruption of the PCP genes Celsr1 and Vangl2, rendering the epithelium almost incapable of branching in response to an FGF10 stimulus.


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)

Mutant lung endoderm responds to FGF10 but is unable to undergo branching. (A, B) Wild-type lung endoderm branches in response to 400 ng/ml FGF10 (mean bud number = 11, n = 6). (C, D) mutant lungs do not branch in response to FGF10 (mean bud number Crsh = 0.9, n = 6). P-ERK1/2 staining of E14.5 lung, shows no significant difference in percentage of P-ERK1/2 positive cells in the epithelium of mutants compared with wild-type controls (E): wild-type 55.17%, ±3, n = 10; Crsh 51.97%, ±6.3, n = 7, P = 0.6074; Lp 56.45%, ±3.9, n = 9, P = 0.7939. Data from three embryos for each genotype. Western blotting reveals no change in the relative levels of P-ERK1/2 in wild-type and Lp/Lp or Crsh/Crsh littermates (F). Scale bars: (A–D) 5 µM.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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

DDQ104F6: Mutant lung endoderm responds to FGF10 but is unable to undergo branching. (A, B) Wild-type lung endoderm branches in response to 400 ng/ml FGF10 (mean bud number = 11, n = 6). (C, D) mutant lungs do not branch in response to FGF10 (mean bud number Crsh = 0.9, n = 6). P-ERK1/2 staining of E14.5 lung, shows no significant difference in percentage of P-ERK1/2 positive cells in the epithelium of mutants compared with wild-type controls (E): wild-type 55.17%, ±3, n = 10; Crsh 51.97%, ±6.3, n = 7, P = 0.6074; Lp 56.45%, ±3.9, n = 9, P = 0.7939. Data from three embryos for each genotype. Western blotting reveals no change in the relative levels of P-ERK1/2 in wild-type and Lp/Lp or Crsh/Crsh littermates (F). Scale bars: (A–D) 5 µM.
Mentions: We wished to determine whether the tissue morphogenesis defects observed in Celsr1 and Vangl2 mutant lungs could reflect a defect in the response of mutant endoderm to a key signal for branching. To test this idea we exposed Celsr1Crsh, Vangl2Lp and wild-type lung epithelium denuded of mesenchyme to FGF10, which normally signals from the mesenchyme to direct lung branching. Both wild-type and mutant lungs responded to the FGF10 stimulus in terms of growth. However, whereas wild-type epithelium formed multiple long and narrow buds (Fig. 6A and B), in most cases, the mutant lung epithelium did not undergo any branching in response to FGF10 (Fig. 6C, D or rarely one or two short stumpy buds formed). Staining for phospho-ERK1/2, which is up-regulated in response to FGF10 in the lung (40), revealed no significant difference in the number of positive cells in wild-type and mutant lung epithelium and this was confirmed by western blot (Fig. 6E and F). Together these results indicate that the FGF signalling pathway, at least that mediated by ERK1/2 activators, is unaffected in mutant lungs. Thus, normal morphogenetic movement within the epithelium is affected upon disruption of the PCP genes Celsr1 and Vangl2, rendering the epithelium almost incapable of branching in response to an FGF10 stimulus.

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