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 in | The Journal of biological chemistry Vol. 268; no. 13; pp. 9645 - 9652 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
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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. |
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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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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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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 |
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