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PMID: 7798324 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Bone morphogenetic protein-2 converts the differentiation pathway of C2C12 myoblasts into the osteoblast lineage.

The Journal of cell biology ·Vol. 127 ·No. 6 Pt 1 ·1994-12-00 ·Pages 1755-66

Katagiri T, Yamaguchi A, Komaki M, Abe E, Takahashi N, Ikeda T, Rosen V, Wozney JM, Fujisawa-Sehara A, Suda T

Abstract

The implantation of bone morphogenetic protein (BMP) into muscular tissues induces ectopic bone formation at the site of implantation. To investigate the mechanism underlying this process, we examined whether recombinant bone morphogenetic protein-2 (BMP-2) converts the differentiation pathway of the clonal myoblastic cell line, C2C12, into that of osteoblast lineage. Incubating the cells with 300 ng/ml of BMP-2 for 6 d almost completely inhibited the formation of the multinucleated myotubes expressing troponin T and myosin heavy chain, and induced the appearance of numerous alkaline phosphatase (ALP)-positive cells. BMP-2 dose dependently induced ALP activity, parathyroid hormone (PTH)-dependent 3',5'-cAMP production, and osteocalcin production at concentrations above 100 ng/ml. The concentration of BMP-2 required to induce these osteoblastic phenotypes was the same as that required to almost completely inhibit myotube formation. Incubating primary muscle cells with 300 ng/ml of BMP-2 for 6 d also inhibited myotube formation, whereas induced ALP activity and osteocalcin production. Incubation with 300 ng/ml of BMP-2 suppressed the expression of mRNA for muscle creatine kinase within 6 h, whereas it induced mRNA expression for ALP, PTH/PTH-related protein (PTHrP) receptors, and osteocalcin within 24-48 h. BMP-2 completely inhibited the expression of myogenin mRNA by day 3. By day 3, BMP-2 also inhibited the expression of MyoD mRNA, but it was transiently stimulated 12 h after exposure to BMP-2. Expression of Id-1 mRNA was greatly stimulated by BMP-2. When C2C12 cells pretreated with BMP-2 for 6 d were transferred to a colony assay system in the absence of BMP-2, more than 84% of the colonies generated became troponin T-positive and ALP activity disappeared. TGF-beta 1 also inhibited myotube formation in C2C12 cells, and suppressed the expression of myogenin and MyoD mRNAs without inducing that of Id-1 mRNA. However, no osteoblastic phenotype was induced by TGF-beta 1 in C2C12 cells. TGF-beta 1 potentiated the inhibitory effect of BMP-2 on myotube formation, whereas TGF-beta 1 reduced ALP activity and osteocalcin production induced by BMP-2 in C2C12 cells. These results indicate that BMP-2 specifically converts the differentiation pathway of C2C12 myoblasts into that of osteoblast lineage cells, but that the conversion is not heritable.

MeSH Terms
Alkaline Phosphatase/biosynthesis Animals Bone Development/physiology Bone Morphogenetic Proteins Cell Differentiation/drug effects Creatine Kinase/biosynthesis Cyclic AMP/biosynthesis DNA-Binding Proteins/biosynthesis,genetics Dose-Response Relationship, Drug Helix-Loop-Helix Motifs Inhibitor of Differentiation Protein 1 Mice Muscles/cytology,drug effects,embryology MyoD Protein/biosynthesis,genetics Myogenin/biosynthesis,genetics Osteoblasts/physiology Osteocalcin/biosynthesis Parathyroid Hormone/biosynthesis Phenotype Proteins/pharmacology RNA, Messenger/analysis Repressor Proteins Stem Cells/drug effects Time Factors Transcription Factors Transforming Growth Factor beta/pharmacology
Chemicals
Bone Morphogenetic Proteins DNA-Binding Proteins Idb1 protein, mouse Inhibitor of Differentiation Protein 1 MyoD Protein Myog protein, mouse Myogenin Parathyroid Hormone Proteins RNA, Messenger Repressor Proteins Transcription Factors Transforming Growth Factor beta Osteocalcin Cyclic AMP Creatine Kinase Alkaline Phosphatase
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Katagiri T
Department of Biochemistry, Showa University, Tokyo, Japan.
Yamaguchi A
Komaki M
Abe E
Takahashi N
Ikeda T
Rosen V
Wozney J M
Fujisawa-Sehara A
Suda T
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.127.6.1755
Published
1994-12-00
Pages
1755-66
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120318
Subset
IM
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1083/jcb.127.6.1755 PMC Full Text View journal details
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