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Altered lung morphogenesis, epithelial cell differentiation and mechanics in mice deficient in the Wnt/β-catenin antagonist Chibby.

Love D, Li FQ, Burke MC, Cyge B, Ohmitsu M, Cabello J, Larson JE, Brody SL, Cohen JC, Takemaru K - PLoS ONE (2010)

Bottom Line: Notably, Cby protein is prominently associated with the centrosome/basal body microtubule structures in embryonic lung epithelial progenitor cells, and later enriches as discrete foci at the base of motile cilia in airway ciliated cells.At birth, Cby(-/-) lungs are grossly normal but spontaneously develop alveolar airspace enlargement with reduced proliferation and abnormal differentiation of lung epithelial cells, resulting in altered pulmonary function.Collectively, our results demonstrate that Cby facilitates proper postnatal lung development and function.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacological Sciences, SUNY at Stony Brook, Stony Brook, New York, United States of America.

ABSTRACT
The canonical Wnt/β-catenin pathway plays crucial roles in various aspects of lung morphogenesis and regeneration/repair. Here, we examined the lung phenotype and function in mice lacking the Wnt/β-catenin antagonist Chibby (Cby). In support of its inhibitory role in canonical Wnt signaling, expression of β-catenin target genes is elevated in the Cby(-/-) lung. Notably, Cby protein is prominently associated with the centrosome/basal body microtubule structures in embryonic lung epithelial progenitor cells, and later enriches as discrete foci at the base of motile cilia in airway ciliated cells. At birth, Cby(-/-) lungs are grossly normal but spontaneously develop alveolar airspace enlargement with reduced proliferation and abnormal differentiation of lung epithelial cells, resulting in altered pulmonary function. Consistent with the Cby expression pattern, airway ciliated cells exhibit a marked paucity of motile cilia with apparent failure of basal body docking. Moreover, we demonstrate that Cby is a direct downstream target for the master ciliogenesis transcription factor Foxj1. Collectively, our results demonstrate that Cby facilitates proper postnatal lung development and function.

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Proximal airway phenotypes of Cby−/− mice.(A) Lung airway sections from adult Cby+/+ and Cby−/− mice were stained with H&E. Motile cilia were noticeable in the airway epithelium of Cby+/+ mice (black arrowheads and inset) but not in that of Cby−/− mice. Atypical morphology of non-ciliated Clara cells was also observed in Cby−/− mice (white arrowheads and inset). Asterisks indicate the airway lumen. (B) Airway sections from adult Cby+/+ and Cby−/− mice were double-labeled with antibodies against the ciliated cell marker acetylated α-tubulin (green) and Clara cell marker CC10 (red), and merged images are shown. Nuclei were stained with DAPI. (C–F) TEM was performed on adult proximal lungs from Cby+/+ (C, E) and Cby−/− (D, F) mice. The airway epithelium of Cby+/+ mice was lined with typical columnar ciliated and non-ciliated Clara cells. Strikingly abnormal morphology and disorganization of these cell types were seen in Cby−/− mice. In addition, ciliated cells had a marked paucity of motile cilia. High-magnification images of ciliated cells revealed that basal bodies (white arrowheads) were polarized perpendicular to the apical cell surface in Cby+/+ mice (E), but frequently misoriented in Cby−/− mice (F). Ci, ciliated cells; CL, Clara cells. Scale bars: (A) 10 µm; (inset) 5 µm; (B) 50 µm; (C, D) 5 µm; (E, F) 500 nm.
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pone-0013600-g007: Proximal airway phenotypes of Cby−/− mice.(A) Lung airway sections from adult Cby+/+ and Cby−/− mice were stained with H&E. Motile cilia were noticeable in the airway epithelium of Cby+/+ mice (black arrowheads and inset) but not in that of Cby−/− mice. Atypical morphology of non-ciliated Clara cells was also observed in Cby−/− mice (white arrowheads and inset). Asterisks indicate the airway lumen. (B) Airway sections from adult Cby+/+ and Cby−/− mice were double-labeled with antibodies against the ciliated cell marker acetylated α-tubulin (green) and Clara cell marker CC10 (red), and merged images are shown. Nuclei were stained with DAPI. (C–F) TEM was performed on adult proximal lungs from Cby+/+ (C, E) and Cby−/− (D, F) mice. The airway epithelium of Cby+/+ mice was lined with typical columnar ciliated and non-ciliated Clara cells. Strikingly abnormal morphology and disorganization of these cell types were seen in Cby−/− mice. In addition, ciliated cells had a marked paucity of motile cilia. High-magnification images of ciliated cells revealed that basal bodies (white arrowheads) were polarized perpendicular to the apical cell surface in Cby+/+ mice (E), but frequently misoriented in Cby−/− mice (F). Ci, ciliated cells; CL, Clara cells. Scale bars: (A) 10 µm; (inset) 5 µm; (B) 50 µm; (C, D) 5 µm; (E, F) 500 nm.

Mentions: In the epithelial lining of the proximal airways, there are two dominant cell types, secretory Clara cells and ciliated cells responsible for mucociliary clearance [29], [30]. The airway epithelium of adult Cby+/+ mice showed a typical pseudostratified columnar morphology with cilia (black arrowheads and inset in Figure 7A). On the contrary, cilia were difficult to detect in Cby−/− mice. In addition, the apical protrusions of Clara cells were more prominent (white arrowheads and inset). The abnormal morphology of the airway epithelial cells was also confirmed by scanning EM (SEM) (Figure S3). Consistent with our histological and SEM data, in the large airway of adult Cby−/− mice, ciliary staining was greatly reduced as shown by immunofluorescent staining for acetylated α-tubulin (Figure 7B). In sharp contrast, there was a dramatic increase in staining for the Clara cell marker CC10 in Cby−/− lungs. It should be noted, however, that we found no evidence of intermediate cell types co-expressing CC10 and the ciliated cell marker Foxj1 in the airway epithelium of adult Cby−/− mice (data not shown).


Altered lung morphogenesis, epithelial cell differentiation and mechanics in mice deficient in the Wnt/β-catenin antagonist Chibby.

Love D, Li FQ, Burke MC, Cyge B, Ohmitsu M, Cabello J, Larson JE, Brody SL, Cohen JC, Takemaru K - PLoS ONE (2010)

Proximal airway phenotypes of Cby−/− mice.(A) Lung airway sections from adult Cby+/+ and Cby−/− mice were stained with H&E. Motile cilia were noticeable in the airway epithelium of Cby+/+ mice (black arrowheads and inset) but not in that of Cby−/− mice. Atypical morphology of non-ciliated Clara cells was also observed in Cby−/− mice (white arrowheads and inset). Asterisks indicate the airway lumen. (B) Airway sections from adult Cby+/+ and Cby−/− mice were double-labeled with antibodies against the ciliated cell marker acetylated α-tubulin (green) and Clara cell marker CC10 (red), and merged images are shown. Nuclei were stained with DAPI. (C–F) TEM was performed on adult proximal lungs from Cby+/+ (C, E) and Cby−/− (D, F) mice. The airway epithelium of Cby+/+ mice was lined with typical columnar ciliated and non-ciliated Clara cells. Strikingly abnormal morphology and disorganization of these cell types were seen in Cby−/− mice. In addition, ciliated cells had a marked paucity of motile cilia. High-magnification images of ciliated cells revealed that basal bodies (white arrowheads) were polarized perpendicular to the apical cell surface in Cby+/+ mice (E), but frequently misoriented in Cby−/− mice (F). Ci, ciliated cells; CL, Clara cells. Scale bars: (A) 10 µm; (inset) 5 µm; (B) 50 µm; (C, D) 5 µm; (E, F) 500 nm.
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Related In: Results  -  Collection

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pone-0013600-g007: Proximal airway phenotypes of Cby−/− mice.(A) Lung airway sections from adult Cby+/+ and Cby−/− mice were stained with H&E. Motile cilia were noticeable in the airway epithelium of Cby+/+ mice (black arrowheads and inset) but not in that of Cby−/− mice. Atypical morphology of non-ciliated Clara cells was also observed in Cby−/− mice (white arrowheads and inset). Asterisks indicate the airway lumen. (B) Airway sections from adult Cby+/+ and Cby−/− mice were double-labeled with antibodies against the ciliated cell marker acetylated α-tubulin (green) and Clara cell marker CC10 (red), and merged images are shown. Nuclei were stained with DAPI. (C–F) TEM was performed on adult proximal lungs from Cby+/+ (C, E) and Cby−/− (D, F) mice. The airway epithelium of Cby+/+ mice was lined with typical columnar ciliated and non-ciliated Clara cells. Strikingly abnormal morphology and disorganization of these cell types were seen in Cby−/− mice. In addition, ciliated cells had a marked paucity of motile cilia. High-magnification images of ciliated cells revealed that basal bodies (white arrowheads) were polarized perpendicular to the apical cell surface in Cby+/+ mice (E), but frequently misoriented in Cby−/− mice (F). Ci, ciliated cells; CL, Clara cells. Scale bars: (A) 10 µm; (inset) 5 µm; (B) 50 µm; (C, D) 5 µm; (E, F) 500 nm.
Mentions: In the epithelial lining of the proximal airways, there are two dominant cell types, secretory Clara cells and ciliated cells responsible for mucociliary clearance [29], [30]. The airway epithelium of adult Cby+/+ mice showed a typical pseudostratified columnar morphology with cilia (black arrowheads and inset in Figure 7A). On the contrary, cilia were difficult to detect in Cby−/− mice. In addition, the apical protrusions of Clara cells were more prominent (white arrowheads and inset). The abnormal morphology of the airway epithelial cells was also confirmed by scanning EM (SEM) (Figure S3). Consistent with our histological and SEM data, in the large airway of adult Cby−/− mice, ciliary staining was greatly reduced as shown by immunofluorescent staining for acetylated α-tubulin (Figure 7B). In sharp contrast, there was a dramatic increase in staining for the Clara cell marker CC10 in Cby−/− lungs. It should be noted, however, that we found no evidence of intermediate cell types co-expressing CC10 and the ciliated cell marker Foxj1 in the airway epithelium of adult Cby−/− mice (data not shown).

Bottom Line: Notably, Cby protein is prominently associated with the centrosome/basal body microtubule structures in embryonic lung epithelial progenitor cells, and later enriches as discrete foci at the base of motile cilia in airway ciliated cells.At birth, Cby(-/-) lungs are grossly normal but spontaneously develop alveolar airspace enlargement with reduced proliferation and abnormal differentiation of lung epithelial cells, resulting in altered pulmonary function.Collectively, our results demonstrate that Cby facilitates proper postnatal lung development and function.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacological Sciences, SUNY at Stony Brook, Stony Brook, New York, United States of America.

ABSTRACT
The canonical Wnt/β-catenin pathway plays crucial roles in various aspects of lung morphogenesis and regeneration/repair. Here, we examined the lung phenotype and function in mice lacking the Wnt/β-catenin antagonist Chibby (Cby). In support of its inhibitory role in canonical Wnt signaling, expression of β-catenin target genes is elevated in the Cby(-/-) lung. Notably, Cby protein is prominently associated with the centrosome/basal body microtubule structures in embryonic lung epithelial progenitor cells, and later enriches as discrete foci at the base of motile cilia in airway ciliated cells. At birth, Cby(-/-) lungs are grossly normal but spontaneously develop alveolar airspace enlargement with reduced proliferation and abnormal differentiation of lung epithelial cells, resulting in altered pulmonary function. Consistent with the Cby expression pattern, airway ciliated cells exhibit a marked paucity of motile cilia with apparent failure of basal body docking. Moreover, we demonstrate that Cby is a direct downstream target for the master ciliogenesis transcription factor Foxj1. Collectively, our results demonstrate that Cby facilitates proper postnatal lung development and function.

Show MeSH
Related in: MedlinePlus