Analysis of type II and type X collagen synthesis in cultured growth plate chondrocytes by in situ hybridization: Rapid induction of type X collagen in culture

2009 ◽  
Vol 9 (11) ◽  
pp. 1713-1722 ◽  
Author(s):  
R.J. O'Keefe ◽  
J.E. Puzas ◽  
L. Loveys ◽  
D.G. Hicks ◽  
R.N. Rosier
Keyword(s):  
In Situ Hybridization ◽  
Growth Plate ◽  
Collagen Synthesis ◽  
Type Ii ◽  
Type X Collagen ◽  
Growth Plate Chondrocytes ◽  
Rapid Induction

scholarly journals In vivo and in vitro Chondrocyte Metabolism in Relationship to the Developemnt of Tibial Dyschondroplasia in Broiler Chickens

1993 ◽  
Author(s):  
Roland M. Leach ◽  
Mark Pines ◽  
Carol V. Gay ◽  
Shmuel Hurwitz
Keyword(s):  
In Situ Hybridization ◽  
Growth Plate ◽  
Bone Growth ◽  
Future Research ◽  
Epiphyseal Growth Plate ◽  
Growth Plate Chondrocytes ◽  
Tibial Dyschondroplasia ◽  
Sequence Of Events

Skeletal deformities are a significant financial and welfare problem for the world poultry industry. Tibial dyschondroplasia (TD) is the most prevalent skeletal abnormality found in young broilers, turkeys and ducks. Tibial dyschondroplasia results from a perturbation of the sequence of events in the epiphyseal growth plate, the tissue responsible for longitudinal bone growth. The purpose of this investigation was to test the hypothesis that TD was the result of a failure of growth plate chondrocytes to differentiate and express the chemotactic molecules required for cartilage vascularization. In this investigation in situ hybridization and immunocytochemical techniques were used to study chondrocyte gene products associated with cartilage maturation and vascularization such as osteopontin, osteonectin, type X collagen, and alkaline phosphatase. All markers were present in the growth plate tissue anter or to the TD lesion but were greatly diminished in the TD lesion. Thus, rather than not acquiring the markers for hypertrophy, it appears that the growth plate chondrocytes reach a certain stage of hypertrophy and then de-differentiate into cells which resemble chondrocytes in the prehypertrophic zone. Similar patterns were observed in all TD tissues examined whether the lesions were spontaneous or induced by dietary treatments or genetic selection. The decrease in gene expression can at least be partially explained by the fact that many of the dysplastic chondrocytes show classic signs of apoptosis. These results provide an explanation for the observation that a variety of genes show reduced expression in the TD lesion when examined by in situ hybridization. This would suggest that future research should focus on the earliest detectable stages in the development of TD and examine endocrine and autocrine factors which cause chondrocytes to de-differentiate and undergo premature apoptosis.

The synthesis of type X collagen by bovine and human growth-plate chondrocytes

1991 ◽  
Vol 99 (3) ◽  
pp. 641-649 ◽  
Author(s):  
A. Marriott ◽  
S. Ayad ◽  
M.E. Grant
Keyword(s):  
Growth Plate ◽  
Type I Collagen ◽  
Polyacrylamide Gel Electrophoresis ◽  
Human Growth ◽  
Type I ◽  
Collagen Gels ◽  
Type X Collagen ◽  
Growth Plate Chondrocytes ◽  
Growth Plate Cartilage ◽  
Disulphide Bonds

Chondrocytes were isolated from bovine growth-plate cartilage and cultured within type I collagen gels. A major collagen with chains of Mr 59,000, decreasing to 47,000 on pepsinization, was synthesized and identified as type X collagen. This collagen was cleaved at two sites by mammalian collagenase, resulting in a major triple-helical fragment with chains of Mr 32,000. The species of Mr 59,000, 47,000 and 32,000 were not detected by SDS-polyacrylamide gel electrophoresis before reduction, indicating the presence of disulphide bonds within the triple helix. In contrast, similar biosynthetic studies with human growth-plate cartilage in organ culture, indicated that human type X collagen does not contain disulphide bonds. A polyclonal antiserum was raised to bovine type X collagen and used in immunolocalization studies to provide direct evidence for the association of type X collagen with the hypertrophic chondrocytes in both bovine and human growth plates during development.

In situ hybridization reveals differential spatial distribution of mRNAs for type I and type II collagen in the chick limb bud

Development ◽  
1988 ◽  
Vol 103 (1) ◽  
pp. 111-118 ◽  
Author(s):  
C.J. Devlin ◽  
P.M. Brickell ◽  
E.R. Taylor ◽  
A. Hornbruch ◽  
R.K. Craig ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Limb Development ◽  
Type I Collagen ◽  
Type Ii Collagen ◽  
Limb Bud ◽  
Type I ◽  
Type Ii ◽  
Mrna Accumulation ◽  
Rna Probes

During limb development, type I collagen disappears from the region where cartilage develops and synthesis of type II collagen, which is characteristic of cartilage, begins. In situ hybridization using antisense RNA probes was used to investigate the spatial localization of type I and type II collagen mRNAs. The distribution of the mRNA for type II collagen corresponded well with the pattern of type II collagen synthesis, suggesting control at the level of transcription and mRNA accumulation. In contrast, the pattern of mRNA for type I collagen remained more or less uniform and did not correspond with the synthesis of the protein, suggesting control primarily at the level of translation or of RNA processing.

Collagen types IX and X in the developing chick tibiotarsus: analyses of mRNAs and proteins

Development ◽  
1991 ◽  
Vol 111 (1) ◽  
pp. 191-196 ◽  
Author(s):  
T.F. Linsenmayer ◽  
Q.A. Chen ◽  
E. Gibney ◽  
M.K. Gordon ◽  
J.K. Marchant ◽  
...  
Keyword(s):  
Translational Regulation ◽  
Type Ii ◽  
Embryonic Chick ◽  
Time Data ◽  
Type X Collagen ◽  
Collagen Types ◽  
Marrow Cavity ◽  
Cartilage Development ◽  
Collagen Protein

To examine the regulation of collagen types IX and X during the hypertrophic phase of endochondral cartilage development, we have employed in situ hybridization and immunofluorescence histochemistry on selected stages of embryonic chick tibiotarsi. The data show that mRNA for type X collagen appears at or about the time that we detect the first appearance of the protein. This result is incompatible with translational regulation, which would require accumulation of the mRNA to occur at an appreciably earlier time. Data on later-stage embryos demonstrate that once hypertrophic chondrocytes initiate synthesis of type X collagen, they sustain high levels of its mRNA during the remainder of the hypertrophic program. This suggests that these cells maintain their integrity until close to the time that they are removed at the advancing marrow cavity. Type X collagen protein in the hypertrophic matrix also extends to the marrow cavity. Type IX collagen is found throughout the hypertrophic matrix, as well as throughout the younger cartilaginous matrices. But the mRNA for this molecule is largely or completely absent from the oldest hypertrophic cells. These data are consistent with a model that we have previously proposed in which newly synthesized type X collagen within the hypertrophic zone can become associated with type II/IX collagen fibrils synthesized and deposited earlier in development (Schmid and Linsenmayer, 1990; Chen et al. 1990).

In situ deformation of growth plate chondrocytes in stress-controlled static vs dynamic compression

2017 ◽  
Vol 56 ◽  
pp. 76-82 ◽  
Author(s):  
Elizabeth A. Zimmermann ◽  
Séréna Bouguerra ◽  
Irene Londoño ◽  
Florina Moldovan ◽  
Carl-Éric Aubin ◽  
...  
Keyword(s):  
Growth Plate ◽  
Dynamic Compression ◽  
Growth Plate Chondrocytes ◽  
In Situ Deformation

In Situ Hybridization Analysis of the Expression of the Type II Collagen Gene in the Developing Chicken Limb Bud

1988 ◽  
Vol 8 (4) ◽  
pp. 277-294 ◽  
Author(s):  
Hyun-Duck Nah ◽  
Barbara J. Rodgers ◽  
William M. Kulyk ◽  
Barbara E. Kream ◽  
Robert A. Kosher ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Type Ii Collagen ◽  
Hybridization Analysis ◽  
Limb Bud ◽  
Collagen Gene ◽  
Type Ii

Bcl-2 rearrangement in patients with chronic hepatitis C associated with essential mixed cryoglobulinemia type II

Blood ◽  
2000 ◽  
Vol 96 (8) ◽  
pp. 2910-2912 ◽  
Author(s):  
Yona Kitay-Cohen ◽  
Aliza Amiel ◽  
Nir Hilzenrat ◽  
Dan Buskila ◽  
Yaffa Ashur ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Chronic Hepatitis ◽  
In Situ Hybridization ◽  
Hepatitis C ◽  
Study Groups ◽  
Mixed Cryoglobulinemia ◽  
Hcv Infection ◽  
Type Ii ◽  
Essential Mixed Cryoglobulinemia

Hepatitis C virus (HCV) infection is found in 80% to 90% of patients with essential mixed cryoglobulinemia (EMC) type II, which is associated with monoclonal IgMk produced by monoclonal B cells. It was investigated whether bcl-2 rearrangement is associated with the clonal B-cell proliferation of EMC induced by hepatitis C. The study groups were composed of 15 patients with HCV and EMC, 12 patients with HCV without EMC, and 7 patients with chronic liver disease (CLD) unrelated to HCV. Fluorescence in situ hybridization with probes was applied to JH and to bcl-2 to study whether JH/bcl-2 translocation was present in these patients. Thirteen of 15 (86%) of patients with HCV-related EMC had the JH/bcl-2 translocation, a significantly higher rate than in HCV patients without EMC (16%; P < .001). Bcl-2 rearrangement was not detected in the patients with CLD not related to HCV. The JH/bcl2 translocation may constitute a pathogenetic link for the development of NHL in patients with HCV infection.

KGF regulates pulmonary epithelial proliferation and surfactant protein gene expression in adult rat lung

2000 ◽  
Vol 279 (6) ◽  
pp. L1146-L1158 ◽  
Author(s):  
Toshiyuki Yano ◽  
Robert J. Mason ◽  
Tianli Pan ◽  
Robin R. Deterding ◽  
Larry D. Nielsen ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Epithelial Cells ◽  
Surfactant Protein ◽  
Surfactant Proteins ◽  
Clara Cell ◽  
Cell Protein ◽  
Mrna Levels ◽  
Type Ii ◽  
Cell Hyperplasia

Keratinocyte growth factor (KGF, FGF-7) is a potent mitogen for epithelial cells. We instilled recombinant human KGF to determine the effects of KGF on alveolar epithelial cells. Left lungs of adult rats were instilled intrabronchially with KGF (5 mg/kg) or normal saline. KGF instillation resulted in epithelial cell hyperplasia, and the alveolar bromodeoxyuridine (BrdU) labeling index peaked at 35% on day 2 after instillation. The mRNA levels for the surfactant proteins (SPs) SP-A, SP-B, and SP-D were increased in whole lung tissue on days 1 and 2 after KGF treatment and then returned to control levels on days 3–7. SP-C mRNA levels were increased on days 2–5 after KGF instillation. However, all surfactant protein mRNAs were reduced in type II cells isolated from rats instilled with KGF 2 or 3 days before isolation. These observations were confirmed by in situ hybridization. Instillation of KGF also increased the amount of SP-A and SP-D in lavage fluid. Transcripts for CC10, the 10-kDa Clara cell protein, were decreased. KGF increases the mRNA for the surfactant proteins per lung because of type II cell hyperplasia, but the mRNA per cell is slightly diminished as measured in isolated cells or estimated by in situ hybridization.

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