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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.


Related in: MedlinePlus

Deletion of IFT80 in the embryonic stage.(A) Line drawing showing the timing of tamoxifen administrations for IFT80 deletion in embryonic stage. Arrows above the line indicate the time of tamoxifen administration to the pregnant females at 14.5, 16.5, and 18.5 days postcoitus. The blue arrow below the line indicates the harvest time. (B) Image of IFT80f/f and Col2α1; IFT80f/f newborn mice exposed to tamoxifen at E14.5, E16.5, and E18.5. Arrows indicate the shortened limbs. (C) Alizarin red and Alcian blue staining of hindlimbs of IFT80f/f and Col2α1; IFT80f/f newborn mice. (D) Western blot analysis of IFT80 expression in the cartilage of IFT80f/f and Col2α1; IFT80f/f mice. IFT80 protein level was normalized to GAPDH (n = 3, *P<0.001, significantly different from the IFT80f/f group). (E) qPCR of IFT80 expression. IFT80 expression level was normalized to GAPDH (n = 3, *P<0.001, significantly different from the IFT80f/f group).
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pone.0130618.g001: Deletion of IFT80 in the embryonic stage.(A) Line drawing showing the timing of tamoxifen administrations for IFT80 deletion in embryonic stage. Arrows above the line indicate the time of tamoxifen administration to the pregnant females at 14.5, 16.5, and 18.5 days postcoitus. The blue arrow below the line indicates the harvest time. (B) Image of IFT80f/f and Col2α1; IFT80f/f newborn mice exposed to tamoxifen at E14.5, E16.5, and E18.5. Arrows indicate the shortened limbs. (C) Alizarin red and Alcian blue staining of hindlimbs of IFT80f/f and Col2α1; IFT80f/f newborn mice. (D) Western blot analysis of IFT80 expression in the cartilage of IFT80f/f and Col2α1; IFT80f/f mice. IFT80 protein level was normalized to GAPDH (n = 3, *P<0.001, significantly different from the IFT80f/f group). (E) qPCR of IFT80 expression. IFT80 expression level was normalized to GAPDH (n = 3, *P<0.001, significantly different from the IFT80f/f group).

Mentions: To study the role of IFT80 in cartilage development in the embryonic stage, IFT80f/f mice were mated with Col2α1; IFT80f/f mice. Pregnant mothers were injected with tamoxifen at the indicated times shown in Fig 1A. Newborn mice were harvested for the phenotype analysis. Col2α1; IFT80f/f mice displayed shorted limbs (Fig 1B). Alizarin Red and Alcian blue staining of hindlimbs confirmed both tibias and femurs from Col2α1; IFT80f/f mice were shorter than those from IFT80f/f mice (Fig 1C). Western blot and qPCR confirmed that IFT80 expression was deleted in the cartilage of Col2α1; IFT80f/f newborn mice (Fig 1D and 1E).


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

Yuan X, Yang S - PLoS ONE (2015)

Deletion of IFT80 in the embryonic stage.(A) Line drawing showing the timing of tamoxifen administrations for IFT80 deletion in embryonic stage. Arrows above the line indicate the time of tamoxifen administration to the pregnant females at 14.5, 16.5, and 18.5 days postcoitus. The blue arrow below the line indicates the harvest time. (B) Image of IFT80f/f and Col2α1; IFT80f/f newborn mice exposed to tamoxifen at E14.5, E16.5, and E18.5. Arrows indicate the shortened limbs. (C) Alizarin red and Alcian blue staining of hindlimbs of IFT80f/f and Col2α1; IFT80f/f newborn mice. (D) Western blot analysis of IFT80 expression in the cartilage of IFT80f/f and Col2α1; IFT80f/f mice. IFT80 protein level was normalized to GAPDH (n = 3, *P<0.001, significantly different from the IFT80f/f group). (E) qPCR of IFT80 expression. IFT80 expression level was normalized to GAPDH (n = 3, *P<0.001, significantly different from the IFT80f/f group).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4476593&req=5

pone.0130618.g001: Deletion of IFT80 in the embryonic stage.(A) Line drawing showing the timing of tamoxifen administrations for IFT80 deletion in embryonic stage. Arrows above the line indicate the time of tamoxifen administration to the pregnant females at 14.5, 16.5, and 18.5 days postcoitus. The blue arrow below the line indicates the harvest time. (B) Image of IFT80f/f and Col2α1; IFT80f/f newborn mice exposed to tamoxifen at E14.5, E16.5, and E18.5. Arrows indicate the shortened limbs. (C) Alizarin red and Alcian blue staining of hindlimbs of IFT80f/f and Col2α1; IFT80f/f newborn mice. (D) Western blot analysis of IFT80 expression in the cartilage of IFT80f/f and Col2α1; IFT80f/f mice. IFT80 protein level was normalized to GAPDH (n = 3, *P<0.001, significantly different from the IFT80f/f group). (E) qPCR of IFT80 expression. IFT80 expression level was normalized to GAPDH (n = 3, *P<0.001, significantly different from the IFT80f/f group).
Mentions: To study the role of IFT80 in cartilage development in the embryonic stage, IFT80f/f mice were mated with Col2α1; IFT80f/f mice. Pregnant mothers were injected with tamoxifen at the indicated times shown in Fig 1A. Newborn mice were harvested for the phenotype analysis. Col2α1; IFT80f/f mice displayed shorted limbs (Fig 1B). Alizarin Red and Alcian blue staining of hindlimbs confirmed both tibias and femurs from Col2α1; IFT80f/f mice were shorter than those from IFT80f/f mice (Fig 1C). Western blot and qPCR confirmed that IFT80 expression was deleted in the cartilage of Col2α1; IFT80f/f newborn mice (Fig 1D and 1E).

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