CARTILAGE-SPECIFIC OVEREXPRESSION OF C-PROPEPTIDE OF TYPE II COLLAGEN AFFECTED MATRIX MINERALIZATION IN TRANSGENIC MICE

2003 ◽  
Vol 07 (03n04) ◽  
pp. 183-189
Author(s):  
Yoshito Matsui ◽  
Ken Nakata ◽  
Eijiro Adachi ◽  
Noriyuki Tsumaki ◽  
Tomoatsu Kimura ◽  
...  
Keyword(s):  
Bone Formation ◽  
Transgenic Mice ◽  
Growth Plate ◽  
Type Ii Collagen ◽  
Whole Body ◽  
Endochondral Bone Formation ◽  
Type Ii ◽  
Endochondral Bone ◽  
Matrix Mineralization ◽  
Growth Plate Cartilage

The C-propeptide of type II collagen (CppII) is cleaved from the procollagen molecule at the time of extracellular secretion from chondrocytes, and was reported to localize in the lower hypertrophic zone of the growth plate cartilage. In the present study, the in vivo role of CppII in the process of endochondral bone formation was investigated by cartilage-specific overexpression of CppII in transgenic mice. Two independent lines of transgenic mice were obtained and they showed mild skeletal dysplasia, as evidenced by morphometric measurement of skeletal bones. Whole body staining revealed delayed mineralization of embryonic endochondral bones, including occipital bone and vertebral bodies. Histological sections showed reduced area of mineralization and scattered chondrocyte hypertrophy in the lower part of growth plate cartilage in the embryonic long bones. Immuno-electron micrographs demonstrated that CppII co-localized with collagen fibrils in the extracellular matrix of the cartilage. Taken together, these results indicate that overexpression of CppII affected endochondral bone formation by negatively regulating the matrix mineralization.

scholarly journals Parathyroid hormone-related peptide-depleted mice show abnormal epiphyseal cartilage development and altered endochondral bone formation.

1994 ◽  
Vol 126 (6) ◽  
pp. 1611-1623 ◽  
Author(s):  
N Amizuka ◽  
H Warshawsky ◽  
J E Henderson ◽  
D Goltzman ◽  
A C Karaplis
Keyword(s):  
Bone Formation ◽  
Embryonic Stem ◽  
Type Ii Collagen ◽  
Epiphyseal Plate ◽  
Epiphyseal Cartilage ◽  
Endochondral Bone Formation ◽  
Type Ii ◽  
Endochondral Bone ◽  
Cartilage Development ◽  
Calcified Cartilage

To elucidate the role of PTHrP in skeletal development, we examined the proximal tibial epiphysis and metaphysis of wild-type (PTHrP-normal) 18-19-d-old fetal mice and of chondrodystrophic litter mates homozygous for a disrupted PTHrP allele generated via homologous recombination in embryonic stem cells (PTHrP-depleted). In the PTHrP-normal epiphysis, immunocytochemistry showed PTHrP to be localized in chondrocytes within the resting zone and at the junction between proliferative and hypertrophic zones. In PTHrP-depleted epiphyses, a diminished [3H]thymidine-labeling index was observed in the resting and proliferative zones accounting for reduced numbers of epiphyseal chondrocytes and for a thinner epiphyseal plate. In the mutant hypertrophic zone, enlarged chondrocytes were interspersed with clusters of cells that did not hypertrophy, but resembled resting or proliferative chondrocytes. Although the overall content of type II collagen in the epiphyseal plate was diminished, the lacunae of these non-hypertrophic chondrocytes did react for type II collagen. Moreover, cell membrane-associated chondroitin sulfate immunoreactivity was evident on these cells. Despite the presence of alkaline phosphatase activity on these nonhypertrophic chondrocytes, the adjacent cartilage matrix did not calcify and their persistence accounted for distorted chondrocyte columns and sporadic distribution of calcified cartilage. Consequently, in the metaphysis, bone deposited on the irregular and sparse scaffold of calcified cartilage and resulted in mixed spicules that did not parallel the longitudinal axis of the tibia and were, therefore, inappropriate for bone elongation. Thus, PTHrP appears to modulate both the proliferation and differentiation of chondrocytes and its absence alters the temporal and spatial sequence of epiphyseal cartilage development and of subsequent endochondral bone formation necessary for normal elongation of long bones.

scholarly journals On the role of type IX collagen in the extracellular matrix of cartilage: type IX collagen is localized to intersections of collagen fibrils.

1986 ◽  
Vol 102 (5) ◽  
pp. 1931-1939 ◽  
Author(s):  
W Müller-Glauser ◽  
B Humbel ◽  
M Glatt ◽  
P Sträuli ◽  
K H Winterhalter ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Bone Formation ◽  
Tissue Distribution ◽  
Chicken Embryo ◽  
Polyclonal Antibodies ◽  
Type Ii Collagen ◽  
Endochondral Bone Formation ◽  
Type Ii ◽  
Endochondral Bone

The tissue distribution of type II and type IX collagen in 17-d-old chicken embryo was studied by immunofluorescence using polyclonal antibodies against type II collagen and a peptic fragment of type IX collagen (HMW), respectively. Both proteins were found only in cartilage where they were co-distributed. They occurred uniformly throughout the extracellular matrix, i.e., without distinction between pericellular, territorial, and interterritorial matrices. Tissues that undergo endochondral bone formation contained type IX collagen, whereas periosteal and membranous bones were negative. The thin collagenous fibrils in cartilage consisted of type II collagen as determined by immunoelectron microscopy. Type IX collagen was associated with the fibrils but essentially was restricted to intersections of the fibrils. These observations suggested that type IX collagen contributes to the stabilization of the network of thin fibers of the extracellular matrix of cartilage by interactions of its triple helical domains with several fibrils at or close to their intersections.

scholarly journals Conditional Deletion of Prolyl Hydroxylase Domain-Containing Protein 2 (Phd2) Gene Reveals Its Essential Role in Chondrocyte Function and Endochondral Bone Formation

Endocrinology ◽  
2016 ◽  
Vol 157 (1) ◽  
pp. 127-140 ◽  
Author(s):  
Shaohong Cheng ◽  
Weirong Xing ◽  
Sheila Pourteymoor ◽  
Jan Schulte ◽  
Subburaman Mohan
Keyword(s):  
Bone Formation ◽  
Growth Plate ◽  
Prolyl Hydroxylase ◽  
Conditional Knockout ◽  
Hypertrophic Chondrocytes ◽  
Endochondral Bone Formation ◽  
Bone Surface ◽  
Endochondral Bone ◽  
Prolyl Hydroxylase Domain

Abstract The hypoxic growth plate cartilage requires hypoxia-inducible factor (HIF)-mediated pathways to maintain chondrocyte survival and differentiation. HIF proteins are tightly regulated by prolyl hydroxylase domain-containing protein 2 (Phd2)-mediated proteosomal degradation. We conditionally disrupted the Phd2 gene in chondrocytes by crossing Phd2 floxed mice with type 2 collagen-α1-Cre transgenic mice and found massive increases (>50%) in the trabecular bone mass of long bones and lumbar vertebra of the Phd2 conditional knockout (cKO) mice caused by significant increases in trabecular number and thickness and reductions in trabecular separation. Cortical thickness and tissue mineral density at the femoral middiaphysis of the cKO mice were also significantly increased. Dynamic histomorphometric analyses revealed increased longitudinal length and osteoid surface per bone surface in the primary spongiosa of the cKO mice, suggesting elevated conversion rate from hypertrophic chondrocytes to mineralized bone matrix as well as increased bone formation in the primary spongiosa. In the secondary spongiosa, bone formation measured by mineralizing surface per bone surface and mineral apposition rate were not changed, but resorption was slightly reduced. Increases in the mRNA levels of SRY (sex determining region Y)-box 9, osterix (Osx), type 2 collagen, aggrecan, alkaline phosphatase, bone sialoprotein, vascular endothelial growth factor, erythropoietin, and glycolytic enzymes in the growth plate of cKO mice were detected by quantitative RT-PCR. Immunohistochemistry revealed an increased HIF-1α protein level in the hypertrophic chondrocytes of cKO mice. Infection of chondrocytes isolated from Phd2 floxed mice with adenoviral Cre resulted in similar gene expression patterns as observed in the cKO growth plate chondrocytes. Our findings indicate that Phd2 suppresses endochondral bone formation, in part, via HIF-dependent mechanisms in mice.

scholarly journals Expression of a Truncated, Kinase-Defective TGF-β Type II Receptor in Mouse Skeletal Tissue Promotes Terminal Chondrocyte Differentiation and Osteoarthritis

1997 ◽  
Vol 139 (2) ◽  
pp. 541-552 ◽  
Author(s):  
Rosa Serra ◽  
Mahlon Johnson ◽  
Ellen H. Filvaroff ◽  
James LaBorde ◽  
Daniel M. Sheehan ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Transgenic Mice ◽  
Growth Plate ◽  
Dominant Negative Mutant ◽  
Type I ◽  
Type Ii ◽  
Type X Collagen ◽  
Skeletal Tissue ◽  
Hypertrophic Cartilage ◽  
Growth Plate Cartilage

Members of the TGF-β superfamily are important regulators of skeletal development. TGF-βs signal through heteromeric type I and type II receptor serine/threonine kinases. When over-expressed, a cytoplasmically truncated type II receptor can compete with the endogenous receptors for complex formation, thereby acting as a dominant-negative mutant (DNIIR). To determine the role of TGF-βs in the development and maintenance of the skeleton, we have generated transgenic mice (MT-DNIIR-4 and -27) that express the DNIIR in skeletal tissue. DNIIR mRNA expression was localized to the periosteum/perichondrium, syno-vium, and articular cartilage. Lower levels of DNIIR mRNA were detected in growth plate cartilage. Transgenic mice frequently showed bifurcation of the xiphoid process and sternum. They also developed progressive skeletal degeneration, resulting by 4 to 8 mo of age in kyphoscoliosis and stiff and torqued joints. The histology of affected joints strongly resembled human osteo-arthritis. The articular surface was replaced by bone or hypertrophic cartilage as judged by the expression of type X collagen, a marker of hypertrophic cartilage normally absent from articular cartilage. The synovium was hyperplastic, and cartilaginous metaplasia was observed in the joint space. We then tested the hypothesis that TGF-β is required for normal differentiation of cartilage in vivo. By 4 and 8 wk of age, the level of type X collagen was increased in growth plate cartilage of transgenic mice relative to wild-type controls. Less proteoglycan staining was detected in the growth plate and articular cartilage matrix of transgenic mice. Mice that express DNIIR in skeletal tissue also demonstrated increased Indian hedgehog (IHH) expression. IHH is a secreted protein that is expressed in chondrocytes that are committed to becoming hypertrophic. It is thought to be involved in a feedback loop that signals through the periosteum/ perichondrium to inhibit cartilage differentiation. The data suggest that TGF-β may be critical for multifaceted maintenance of synovial joints. Loss of responsiveness to TGF-β promotes chondrocyte terminal differentiation and results in development of degenerative joint disease resembling osteoarthritis in humans.

scholarly journals Growth plate expression profiling: Large and small breed dogs provide new insights in endochondral bone formation

2017 ◽  
Author(s):  
Michelle Teunissen ◽  
Frank M. Riemers ◽  
Dik van Leenen ◽  
Marian J. A. Groot Koerkamp ◽  
Björn P. Meij ◽  
...  
Keyword(s):  
Bone Formation ◽  
Growth Plate ◽  
Expression Profiling ◽  
Endochondral Bone Formation ◽  
Endochondral Bone ◽  
Small Breed

scholarly journals Galectin-3 Is a Downstream Regulator of Matrix Metalloproteinase-9 Function during Endochondral Bone Formation

2005 ◽  
Vol 16 (6) ◽  
pp. 3028-3039 ◽  
Author(s):  
Nathalie Ortega ◽  
Danielle J. Behonick ◽  
Céline Colnot ◽  
Douglas N.W. Cooper ◽  
Zena Werb
Keyword(s):  
Bone Formation ◽  
Matrix Metalloproteinase ◽  
Growth Plate ◽  
Matrix Metalloproteinase 9 ◽  
Hypertrophic Chondrocytes ◽  
Endochondral Bone Formation ◽  
Endochondral Bone ◽  
Chondrocyte Death ◽  
Galectin 3 ◽  
Metalloproteinase 9

Endochondral bone formation is characterized by the progressive replacement of a cartilage anlagen by bone at the growth plate with a tight balance between the rates of chondrocyte proliferation, differentiation, and cell death. Deficiency of matrix metalloproteinase-9 (MMP-9) leads to an accumulation of late hypertrophic chondrocytes. We found that galectin-3, an in vitro substrate of MMP-9, accumulates in the late hypertrophic chondrocytes and their surrounding extracellular matrix in the expanded hypertrophic cartilage zone. Treatment of wild-type embryonic metatarsals in culture with full-length galectin-3, but not galectin-3 cleaved by MMP-9, mimicked the embryonic phenotype of Mmp-9 null mice, with an increased hypertrophic zone and decreased osteoclast recruitment. These results indicate that extracellular galectin-3 could be an endogenous substrate of MMP-9 that acts downstream to regulate hypertrophic chondrocyte death and osteoclast recruitment during endochondral bone formation. Thus, the disruption of growth plate homeostasis in Mmp-9 null mice links galectin-3 and MMP-9 in the regulation of the clearance of late chondrocytes through regulation of their terminal differentiation.

scholarly journals Association of an extracellular protein (chondrocalcin) with the calcification of cartilage in endochondral bone formation.

1984 ◽  
Vol 98 (1) ◽  
pp. 54-65 ◽  
Author(s):  
A R Poole ◽  
I Pidoux ◽  
A Reiner ◽  
H Choi ◽  
L C Rosenberg
Keyword(s):  
Extracellular Matrix ◽  
Bone Formation ◽  
Growth Plate ◽  
Calcium Binding ◽  
Calcium Binding Protein ◽  
Cartilage Matrix ◽  
Hypertrophic Chondrocytes ◽  
Endochondral Bone Formation ◽  
Endochondral Bone ◽  
Nucleation Sites

We examined bovine fetal epiphyseal and growth plate cartilages by immunofluorescence microscopy and immunoelectron microscopy using monospecific antibodies to a newly discovered cartilage-matrix calcium-binding protein that we now call chondrocalcin. Chondrocalcin was evenly distributed at relatively low concentration in resting fetal epiphyseal cartilage. In growth plate cartilage, it was absent from the extracellular matrix in the zone of proliferating chondrocytes but was present in intracellular vacuoles in proliferating, maturing and upper hypertrophic chondrocytes. The protein then disappeared from the lower hypertrophic chondrocytes and appeared in the adjoining extracellular matrix, where it was selectively concentrated in the longitudinal septa in precisely the same location where amorphous mineral was deposited in large amounts as demonstrated by von Kossa staining and electron microscopy. Mineral then spread out from these "nucleation sites" to occupy much of the surrounding matrix. Matrix vesicles were identified in this calcifying matrix but they bore no observable morphological relationship to these major sites of calcification where chondrocalcin was concentrated. Since chondrocalcin is a calcium-binding protein and has a strong affinity for hydroxyapatite, these observations suggest that chondrocalcin may play a fundamental role in the creation of nucleation sites for the calcification of cartilage matrix in endochondral bone formation.

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