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Deletion of IFT80 Impairs Epiphyseal and Articular Cartilage Formation Due to Disruption of Chondrocyte Differentiation.

Yuan X, Yang S - PLoS ONE (2015)

Bottom Line: Intraflagellar transport proteins (IFT) play important roles in cilia formation and organ development.Loss of IFT80 in the embryonic stage resulted in short limbs at birth.These findings demonstrate that loss of IFT80 blocks chondrocyte differentiation by disruption of ciliogenesis and alteration of Hh and Wnt signaling transduction, which in turn alters epiphyseal and articular cartilage formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Biology, School of Dental Medicine, University of Buffalo, State University of New York, Buffalo, NY, United States of America.

ABSTRACT
Intraflagellar transport proteins (IFT) play important roles in cilia formation and organ development. Partial loss of IFT80 function leads Jeune asphyxiating thoracic dystrophy (JATD) or short-rib polydactyly (SRP) syndrome type III, displaying narrow thoracic cavity and multiple cartilage anomalies. However, it is unknown how IFT80 regulates cartilage formation. To define the role and mechanism of IFT80 in chondrocyte function and cartilage formation, we generated a Col2α1; IFT80f/f mouse model by crossing IFT80f/f mice with inducible Col2α1-CreER mice, and deleted IFT80 in chondrocyte lineage by injection of tamoxifen into the mice in embryonic or postnatal stage. Loss of IFT80 in the embryonic stage resulted in short limbs at birth. Histological studies showed that IFT80-deficient mice have shortened cartilage with marked changes in cellular morphology and organization in the resting, proliferative, pre-hypertrophic, and hypertrophic zones. Moreover, deletion of IFT80 in the postnatal stage led to mouse stunted growth with shortened growth plate but thickened articular cartilage. Defects of ciliogenesis were found in the cartilage of IFT80-deficient mice and primary IFT80-deficient chondrocytes. Further study showed that chondrogenic differentiation was significantly inhibited in IFT80-deficient mice due to reduced hedgehog (Hh) signaling and increased Wnt signaling activities. These findings demonstrate that loss of IFT80 blocks chondrocyte differentiation by disruption of ciliogenesis and alteration of Hh and Wnt signaling transduction, which in turn alters epiphyseal and articular cartilage formation.

No MeSH data available.


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Significant cilia formation defects in the growth plate of Col2α1; IFT80f/f mice.(A) Immunofluorescence analysis of primary cilia in the tibial growth plate of newborn mice exposed to tamoxifen at E14.5, E16.5, and E18.5 (n = 3). Immunostaining of primary cilia was performed with acetylated α-tubulin (axoneme, cyan) antibody. DAPI (nuclear marker) staining was used as counterstain. Scale bars represent 100 μm. (B) Quantification of the ciliated cell population in resting zone, proliferation zone and hypertrophic zone (n = 3, *p<0.0001, significantly different from the IFT80f/f group). (C) Immunofluorescence analysis of primary cilia in the cartilage of P30 mice exposed to tamoxifen from P4-P7 and P14-P17 (n = 3). (D) Quantification of the ciliated cell population in epiphyseal plate and articular cartilage (n = 3, *p<0.0001, significantly different from the IFT80f/f group).
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pone.0130618.g006: Significant cilia formation defects in the growth plate of Col2α1; IFT80f/f mice.(A) Immunofluorescence analysis of primary cilia in the tibial growth plate of newborn mice exposed to tamoxifen at E14.5, E16.5, and E18.5 (n = 3). Immunostaining of primary cilia was performed with acetylated α-tubulin (axoneme, cyan) antibody. DAPI (nuclear marker) staining was used as counterstain. Scale bars represent 100 μm. (B) Quantification of the ciliated cell population in resting zone, proliferation zone and hypertrophic zone (n = 3, *p<0.0001, significantly different from the IFT80f/f group). (C) Immunofluorescence analysis of primary cilia in the cartilage of P30 mice exposed to tamoxifen from P4-P7 and P14-P17 (n = 3). (D) Quantification of the ciliated cell population in epiphyseal plate and articular cartilage (n = 3, *p<0.0001, significantly different from the IFT80f/f group).

Mentions: To further get insight into the mechanism of IFT80 in chondrocyte differentiation, we first performed cilia staining in the tibia section of the newborn mice, which were administered tamoxifen at 14.5, 16.5, and 18.5 days postcoitus. As shown in Fig 6A and 6B, about 80% chondrocytes in the resting zone and proliferation zone, and 70% in hypertrophic zone had normal cilia structure in IFT80f/f mice group. However, only about 20% chondrocytes in those zones showed cilia in Col2α1; IFT80f/f mice group (Fig 6A and 6B). Consistent to the results from the newborn mice, in IFT80f/f mice at postnatal day 30, 40% chondrocytes in epiphyseal plates and 60% in articular cartilage had cilia (Fig 6C and 6D). In contrast, cilia were absent in the growth plates and only present in 10% articular cartilage cells in Col2α1; IFT80f/f mice (Fig 6C and 6D).


Deletion of IFT80 Impairs Epiphyseal and Articular Cartilage Formation Due to Disruption of Chondrocyte Differentiation.

Yuan X, Yang S - PLoS ONE (2015)

Significant cilia formation defects in the growth plate of Col2α1; IFT80f/f mice.(A) Immunofluorescence analysis of primary cilia in the tibial growth plate of newborn mice exposed to tamoxifen at E14.5, E16.5, and E18.5 (n = 3). Immunostaining of primary cilia was performed with acetylated α-tubulin (axoneme, cyan) antibody. DAPI (nuclear marker) staining was used as counterstain. Scale bars represent 100 μm. (B) Quantification of the ciliated cell population in resting zone, proliferation zone and hypertrophic zone (n = 3, *p<0.0001, significantly different from the IFT80f/f group). (C) Immunofluorescence analysis of primary cilia in the cartilage of P30 mice exposed to tamoxifen from P4-P7 and P14-P17 (n = 3). (D) Quantification of the ciliated cell population in epiphyseal plate and articular cartilage (n = 3, *p<0.0001, significantly different from the IFT80f/f group).
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pone.0130618.g006: Significant cilia formation defects in the growth plate of Col2α1; IFT80f/f mice.(A) Immunofluorescence analysis of primary cilia in the tibial growth plate of newborn mice exposed to tamoxifen at E14.5, E16.5, and E18.5 (n = 3). Immunostaining of primary cilia was performed with acetylated α-tubulin (axoneme, cyan) antibody. DAPI (nuclear marker) staining was used as counterstain. Scale bars represent 100 μm. (B) Quantification of the ciliated cell population in resting zone, proliferation zone and hypertrophic zone (n = 3, *p<0.0001, significantly different from the IFT80f/f group). (C) Immunofluorescence analysis of primary cilia in the cartilage of P30 mice exposed to tamoxifen from P4-P7 and P14-P17 (n = 3). (D) Quantification of the ciliated cell population in epiphyseal plate and articular cartilage (n = 3, *p<0.0001, significantly different from the IFT80f/f group).
Mentions: To further get insight into the mechanism of IFT80 in chondrocyte differentiation, we first performed cilia staining in the tibia section of the newborn mice, which were administered tamoxifen at 14.5, 16.5, and 18.5 days postcoitus. As shown in Fig 6A and 6B, about 80% chondrocytes in the resting zone and proliferation zone, and 70% in hypertrophic zone had normal cilia structure in IFT80f/f mice group. However, only about 20% chondrocytes in those zones showed cilia in Col2α1; IFT80f/f mice group (Fig 6A and 6B). Consistent to the results from the newborn mice, in IFT80f/f mice at postnatal day 30, 40% chondrocytes in epiphyseal plates and 60% in articular cartilage had cilia (Fig 6C and 6D). In contrast, cilia were absent in the growth plates and only present in 10% articular cartilage cells in Col2α1; IFT80f/f mice (Fig 6C and 6D).

Bottom Line: Intraflagellar transport proteins (IFT) play important roles in cilia formation and organ development.Loss of IFT80 in the embryonic stage resulted in short limbs at birth.These findings demonstrate that loss of IFT80 blocks chondrocyte differentiation by disruption of ciliogenesis and alteration of Hh and Wnt signaling transduction, which in turn alters epiphyseal and articular cartilage formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Biology, School of Dental Medicine, University of Buffalo, State University of New York, Buffalo, NY, United States of America.

ABSTRACT
Intraflagellar transport proteins (IFT) play important roles in cilia formation and organ development. Partial loss of IFT80 function leads Jeune asphyxiating thoracic dystrophy (JATD) or short-rib polydactyly (SRP) syndrome type III, displaying narrow thoracic cavity and multiple cartilage anomalies. However, it is unknown how IFT80 regulates cartilage formation. To define the role and mechanism of IFT80 in chondrocyte function and cartilage formation, we generated a Col2α1; IFT80f/f mouse model by crossing IFT80f/f mice with inducible Col2α1-CreER mice, and deleted IFT80 in chondrocyte lineage by injection of tamoxifen into the mice in embryonic or postnatal stage. Loss of IFT80 in the embryonic stage resulted in short limbs at birth. Histological studies showed that IFT80-deficient mice have shortened cartilage with marked changes in cellular morphology and organization in the resting, proliferative, pre-hypertrophic, and hypertrophic zones. Moreover, deletion of IFT80 in the postnatal stage led to mouse stunted growth with shortened growth plate but thickened articular cartilage. Defects of ciliogenesis were found in the cartilage of IFT80-deficient mice and primary IFT80-deficient chondrocytes. Further study showed that chondrogenic differentiation was significantly inhibited in IFT80-deficient mice due to reduced hedgehog (Hh) signaling and increased Wnt signaling activities. These findings demonstrate that loss of IFT80 blocks chondrocyte differentiation by disruption of ciliogenesis and alteration of Hh and Wnt signaling transduction, which in turn alters epiphyseal and articular cartilage formation.

No MeSH data available.


Related in: MedlinePlus