Expression and role of c-myc in chondrocytes undergoing endochondral ossification

To analyze the relationship between c-myc gene expression and chondrocyte proliferation and maturation during endochondral ossification, Day 18-19 chick embryo sterna were pulse-labeled with [3H]thymidine, and serial sections were processed for autoradiography and in situ hybridization. Proliferatin...

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Published inThe Journal of biological chemistry Vol. 268; no. 13; pp. 9645 - 9652
Main Authors IWAMOTO, M, YAGAMI, K, VALLE, P. L, OLSEN, B. R, PETROPOULOS, C. J, EWERT, D. L, PACIFICI, M
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Published Bethesda, MD American Society for Biochemistry and Molecular Biology 05.05.1993
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Abstract To analyze the relationship between c-myc gene expression and chondrocyte proliferation and maturation during endochondral ossification, Day 18-19 chick embryo sterna were pulse-labeled with [3H]thymidine, and serial sections were processed for autoradiography and in situ hybridization. Proliferating chondrocytes, located in four distinct areas of the developing sternum, all contained high levels of c-myc transcripts, whereas postmitotic chondrocytes (such as hypertrophic chondrocytes) contained undetectable amounts. These findings were confirmed by Northern blot analysis and by the observation that antisense c-myc oligomer treatment inhibited proliferation in cultured chondrocytes. Constitutive overexpression of c-myc by retroviral vectors in immature chondrocyte cultures (c-myc cultures) maintained the cells in a proliferative state and blocked their maturation into hypertrophic chondrocytes. The lack of maturation in the c-myc cultures was corroborated by analysis of type X collagen gene regulation. Control immature cultures contained strong repressor activity for the type X collagen gene promoter, as revealed by transfection assays; repressor activity was lost upon maturation and activation of type X collagen synthesis. In the c-myc cultures, however, repressor activity persisted. Thus, c-myc participates in the normal changes in proliferation accompanying chondrocyte maturation in vivo and in culture. The decreases in c-myc expression and cell proliferation appear to be required for completion of maturation.
AbstractList To analyze the relationship between c-myc gene expression and chondrocyte proliferation and maturation during endochondral ossification, Day 18-19 chick embryo sterna were pulse-labeled with [3H]thymidine, and serial sections were processed for autoradiography and in situ hybridization. Proliferating chondrocytes, located in four distinct areas of the developing sternum, all contained high levels of c-myc transcripts, whereas postmitotic chondrocytes (such as hypertrophic chondrocytes) contained undetectable amounts. These findings were confirmed by Northern blot analysis and by the observation that antisense c-myc oligomer treatment inhibited proliferation in cultured chondrocytes. Constitutive overexpression of c-myc by retroviral vectors in immature chondrocyte cultures (c-myc cultures) maintained the cells in a proliferative state and blocked their maturation into hypertrophic chondrocytes. The lack of maturation in the c-myc cultures was corroborated by analysis of type X collagen gene regulation. Control immature cultures contained strong repressor activity for the type X collagen gene promoter, as revealed by transfection assays; repressor activity was lost upon maturation and activation of type X collagen synthesis. In the c-myc cultures, however, repressor activity persisted. Thus, c-myc participates in the normal changes in proliferation accompanying chondrocyte maturation in vivo and in culture. The decreases in c-myc expression and cell proliferation appear to be required for completion of maturation.
To analyze the relationship between c-myc gene expression and chondrocyte proliferation and maturation during endochondral ossification, Day 18-19 chick embryo sterna were pulse-labeled with [3H]thymidine, and serial sections were processed for autoradiography and in situ hybridization. Proliferating chondrocytes, located in four distinct areas of the developing sternum, all contained high levels of c-myc transcripts, whereas postmitotic chondrocytes (such as hypertrophic chondrocytes) contained undetectable amounts. These findings were confirmed by Northern blot analysis and by the observation that antisense c-myc oligomer treatment inhibited proliferation in cultured chondrocytes. Constitutive overexpression of c-myc by retroviral vectors in immature chondrocyte cultures (c-myc cultures) maintained the cells in a proliferative state and blocked their maturation into hypertrophic chondrocytes. The lack of maturation in the c-myc cultures was corroborated by analysis of type X collagen gene regulation. Control immature cultures contained strong repressor activity for the type X collagen gene promoter, as revealed by transfection assays; repressor activity was lost upon maturation and activation of type X collagen synthesis. In the c-myc cultures, however, repressor activity persisted. Thus, c-myc participates in the normal changes in proliferation accompanying chondrocyte maturation in vivo and in culture. The decreases in c-myc expression and cell proliferation appear to be required for completion of maturation.
Author M Iwamoto
B R Olsen
K Yagami
M Pacifici
C J Petropoulos
P Lu Valle
D L Ewert
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Issue 13
Keywords Cell proliferation
Vertebrata
Chondrocyte
Collagen
Chicken
Aves
Cell differentiation
Gene expression
Ossification
Language English
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Snippet To analyze the relationship between c-myc gene expression and chondrocyte proliferation and maturation during endochondral ossification, Day 18-19 chick embryo...
To analyze the relationship between c-myc gene expression and chondrocyte proliferation and maturation during endochondral ossification, Day 18-19 chick embryo...
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StartPage 9645
SubjectTerms Animals
Autoradiography
Base Sequence
Biological and medical sciences
Blotting, Northern
Cartilage - embryology
Cartilage - physiology
Cell differentiation, maturation, development, hematopoiesis
Cell Division - drug effects
Cell physiology
Cells, Cultured
Chick Embryo
Collagen - genetics
DNA - biosynthesis
DNA - genetics
DNA Replication
Exons
Fundamental and applied biological sciences. Psychology
Gene Expression
Genes, myc
In Situ Hybridization
Kinetics
Molecular and cellular biology
Molecular Sequence Data
Oligodeoxyribonucleotides
Oligonucleotides, Antisense - pharmacology
Osteogenesis
Promoter Regions, Genetic
RNA, Messenger - genetics
RNA, Messenger - metabolism
Sternum
Thionucleotides - pharmacology
Thymidine - metabolism
Transcription, Genetic
Tritium
Title Expression and role of c-myc in chondrocytes undergoing endochondral ossification
URI http://www.jbc.org/content/268/13/9645.abstract
https://www.ncbi.nlm.nih.gov/pubmed/8486652
Volume 268
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