Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Jan 1;132(1):195–210. doi: 10.1083/jcb.132.1.195

Increased expression of TGF-beta 2 in osteoblasts results in an osteoporosis-like phenotype

PMCID: PMC2120709  PMID: 8567723

Abstract

The development of the skeleton requires the coordinated activities of bone-forming osteoblasts and bone-resorbing osteoclasts. The activities of these two cell types are likely to be regulated by TGF-beta, which is abundant in bone matrix. We have used transgenic mice to evaluate the role of TGF-beta 2 in bone development and turnover. Osteoblast- specific overexpression of TGF-beta 2 from the osteocalcin promoter resulted in progressive bone loss associated with increases in osteoblastic matrix deposition and osteoclastic bone resorption. This phenotype closely resembles the bone abnormalities seen in human hyperparathyroidism and osteoporosis. Furthermore, a high level of TGF- beta 2 overexpression resulted in defective bone mineralization and severe hypoplasia of the clavicles, a hallmark of the developmental disease cleidocranial dysplasia. Our results suggest that TGF-beta 2 functions as a local positive regulator of bone remodeling and that alterations in TGF-beta 2 synthesis by bone cells, or in their responsiveness to TGF-beta 2, may contribute to the pathogenesis of metabolic bone disease.

Full Text

The Full Text of this article is available as a PDF (14.4 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Araki N., Robinson F. D., Nishimoto S. K. Rapid and sensitive method of quantitation of bone gla protein mRNA using competitive polymerase chain reaction. J Bone Miner Res. 1993 Mar;8(3):313–322. doi: 10.1002/jbmr.5650080308. [DOI] [PubMed] [Google Scholar]
  2. Baker A. R., Hollingshead P. G., Pitts-Meek S., Hansen S., Taylor R., Stewart T. A. Osteoblast-specific expression of growth hormone stimulates bone growth in transgenic mice. Mol Cell Biol. 1992 Dec;12(12):5541–5547. doi: 10.1128/mcb.12.12.5541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bonewald L. F., Dallas S. L. Role of active and latent transforming growth factor beta in bone formation. J Cell Biochem. 1994 Jul;55(3):350–357. doi: 10.1002/jcb.240550312. [DOI] [PubMed] [Google Scholar]
  4. Bronckers A. L., Gay S., Dimuzio M. T., Butler W. T. Immunolocalization of gamma-carboxyglutamic acid containing proteins in developing rat bones. Coll Relat Res. 1985 Jun;5(3):273–281. doi: 10.1016/s0174-173x(85)80017-0. [DOI] [PubMed] [Google Scholar]
  5. Brunner A. M., Marquardt H., Malacko A. R., Lioubin M. N., Purchio A. F. Site-directed mutagenesis of cysteine residues in the pro region of the transforming growth factor beta 1 precursor. Expression and characterization of mutant proteins. J Biol Chem. 1989 Aug 15;264(23):13660–13664. [PubMed] [Google Scholar]
  6. Centrella M., Horowitz M. C., Wozney J. M., McCarthy T. L. Transforming growth factor-beta gene family members and bone. Endocr Rev. 1994 Feb;15(1):27–39. doi: 10.1210/edrv-15-1-27. [DOI] [PubMed] [Google Scholar]
  7. Centrella M., McCarthy T. L., Canalis E. Parathyroid hormone modulates transforming growth factor beta activity and binding in osteoblast-enriched cell cultures from fetal rat parietal bone. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5889–5893. doi: 10.1073/pnas.85.16.5889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chenu C., Pfeilschifter J., Mundy G. R., Roodman G. D. Transforming growth factor beta inhibits formation of osteoclast-like cells in long-term human marrow cultures. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5683–5687. doi: 10.1073/pnas.85.15.5683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dieudonné S. C., Foo P., van Zoelen E. J., Burger E. H. Inhibiting and stimulating effects of TGF-beta 1 on osteoclastic bone resorption in fetal mouse bone organ cultures. J Bone Miner Res. 1991 May;6(5):479–487. doi: 10.1002/jbmr.5650060509. [DOI] [PubMed] [Google Scholar]
  10. Engler-Blum G., Meier M., Frank J., Müller G. A. Reduction of background problems in nonradioactive northern and Southern blot analyses enables higher sensitivity than 32P-based hybridizations. Anal Biochem. 1993 May 1;210(2):235–244. doi: 10.1006/abio.1993.1189. [DOI] [PubMed] [Google Scholar]
  11. Erlebacher A., Filvaroff E. H., Gitelman S. E., Derynck R. Toward a molecular understanding of skeletal development. Cell. 1995 Feb 10;80(3):371–378. doi: 10.1016/0092-8674(95)90487-5. [DOI] [PubMed] [Google Scholar]
  12. Finkelman R. D., Bell N. H., Strong D. D., Demers L. M., Baylink D. J. Ovariectomy selectively reduces the concentration of transforming growth factor beta in rat bone: implications for estrogen deficiency-associated bone loss. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):12190–12193. doi: 10.1073/pnas.89.24.12190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Finkelman R. D., Linkhart T. A., Mohan S., Lau K. H., Baylink D. J., Bell N. H. Vitamin D deficiency causes a selective reduction in deposition of transforming growth factor beta in rat bone: possible mechanism for impaired osteoinduction. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3657–3660. doi: 10.1073/pnas.88.9.3657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Groot C. G., Danes J. K., Blok J., Hoogendijk A., Hauschka P. V. Light and electron microscopic demonstration of osteocalcin antigenicity in embryonic and adult rat bone. Bone. 1986;7(5):379–385. doi: 10.1016/8756-3282(86)90259-0. [DOI] [PubMed] [Google Scholar]
  15. Hall B. K. The role of movement and tissue interactions in the development and growth of bone and secondary cartilage in the clavicle of the embryonic chick. J Embryol Exp Morphol. 1986 Apr;93:133–152. [PubMed] [Google Scholar]
  16. Hattersley G., Chambers T. J. Effects of transforming growth factor beta 1 on the regulation of osteoclastic development and function. J Bone Miner Res. 1991 Feb;6(2):165–172. doi: 10.1002/jbmr.5650060210. [DOI] [PubMed] [Google Scholar]
  17. Huang S. S., O'Grady P., Huang J. S. Human transforming growth factor beta.alpha 2-macroglobulin complex is a latent form of transforming growth factor beta. J Biol Chem. 1988 Jan 25;263(3):1535–1541. [PubMed] [Google Scholar]
  18. Joyce M. E., Roberts A. B., Sporn M. B., Bolander M. E. Transforming growth factor-beta and the initiation of chondrogenesis and osteogenesis in the rat femur. J Cell Biol. 1990 Jun;110(6):2195–2207. doi: 10.1083/jcb.110.6.2195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Laird P. W., Zijderveld A., Linders K., Rudnicki M. A., Jaenisch R., Berns A. Simplified mammalian DNA isolation procedure. Nucleic Acids Res. 1991 Aug 11;19(15):4293–4293. doi: 10.1093/nar/19.15.4293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lawrence D. A., Pircher R., Jullien P. Conversion of a high molecular weight latent beta-TGF from chicken embryo fibroblasts into a low molecular weight active beta-TGF under acidic conditions. Biochem Biophys Res Commun. 1985 Dec 31;133(3):1026–1034. doi: 10.1016/0006-291x(85)91239-2. [DOI] [PubMed] [Google Scholar]
  21. Lewis D. B., Liggitt H. D., Effmann E. L., Motley S. T., Teitelbaum S. L., Jepsen K. J., Goldstein S. A., Bonadio J., Carpenter J., Perlmutter R. M. Osteoporosis induced in mice by overproduction of interleukin 4. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11618–11622. doi: 10.1073/pnas.90.24.11618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Manolagas S. C., Jilka R. L. Bone marrow, cytokines, and bone remodeling. Emerging insights into the pathophysiology of osteoporosis. N Engl J Med. 1995 Feb 2;332(5):305–311. doi: 10.1056/NEJM199502023320506. [DOI] [PubMed] [Google Scholar]
  23. Massagué J., Cheifetz S., Endo T., Nadal-Ginard B. Type beta transforming growth factor is an inhibitor of myogenic differentiation. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8206–8210. doi: 10.1073/pnas.83.21.8206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McSheehy P. M., Chambers T. J. Osteoblastic cells mediate osteoclastic responsiveness to parathyroid hormone. Endocrinology. 1986 Feb;118(2):824–828. doi: 10.1210/endo-118-2-824. [DOI] [PubMed] [Google Scholar]
  25. Miller D. A., Lee A., Pelton R. W., Chen E. Y., Moses H. L., Derynck R. Murine transforming growth factor-beta 2 cDNA sequence and expression in adult tissues and embryos. Mol Endocrinol. 1989 Jul;3(7):1108–1114. doi: 10.1210/mend-3-7-1108. [DOI] [PubMed] [Google Scholar]
  26. Noda M., Camilliere J. J. In vivo stimulation of bone formation by transforming growth factor-beta. Endocrinology. 1989 Jun;124(6):2991–2994. doi: 10.1210/endo-124-6-2991. [DOI] [PubMed] [Google Scholar]
  27. O'Connor-McCourt M. D., Wakefield L. M. Latent transforming growth factor-beta in serum. A specific complex with alpha 2-macroglobulin. J Biol Chem. 1987 Oct 15;262(29):14090–14099. [PubMed] [Google Scholar]
  28. O'Hare K., Benoist C., Breathnach R. Transformation of mouse fibroblasts to methotrexate resistance by a recombinant plasmid expressing a prokaryotic dihydrofolate reductase. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1527–1531. doi: 10.1073/pnas.78.3.1527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Olson E. N., Sternberg E., Hu J. S., Spizz G., Wilcox C. Regulation of myogenic differentiation by type beta transforming growth factor. J Cell Biol. 1986 Nov;103(5):1799–1805. doi: 10.1083/jcb.103.5.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Oursler M. J., Cortese C., Keeting P., Anderson M. A., Bonde S. K., Riggs B. L., Spelsberg T. C. Modulation of transforming growth factor-beta production in normal human osteoblast-like cells by 17 beta-estradiol and parathyroid hormone. Endocrinology. 1991 Dec;129(6):3313–3320. doi: 10.1210/endo-129-6-3313. [DOI] [PubMed] [Google Scholar]
  31. PAI A. C. DEVELOPMENTAL GENETICS OF A LETHAL MUTATION, MUSCULAR DYSGENESIS (MDG), IN THE MOUSE. I. GENETIC ANALYSIS AND GROSS MORPHOLOGY. Dev Biol. 1965 Feb;11:82–92. doi: 10.1016/0012-1606(65)90038-2. [DOI] [PubMed] [Google Scholar]
  32. Parfitt A. M. Osteonal and hemi-osteonal remodeling: the spatial and temporal framework for signal traffic in adult human bone. J Cell Biochem. 1994 Jul;55(3):273–286. doi: 10.1002/jcb.240550303. [DOI] [PubMed] [Google Scholar]
  33. Pelton R. W., Saxena B., Jones M., Moses H. L., Gold L. I. Immunohistochemical localization of TGF beta 1, TGF beta 2, and TGF beta 3 in the mouse embryo: expression patterns suggest multiple roles during embryonic development. J Cell Biol. 1991 Nov;115(4):1091–1105. doi: 10.1083/jcb.115.4.1091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pfeilschifter J., Laukhuf F., Müller-Beckmann B., Blum W. F., Pfister T., Ziegler R. Parathyroid hormone increases the concentration of insulin-like growth factor-I and transforming growth factor beta 1 in rat bone. J Clin Invest. 1995 Aug;96(2):767–774. doi: 10.1172/JCI118121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pfeilschifter J., Mundy G. R. Modulation of type beta transforming growth factor activity in bone cultures by osteotropic hormones. Proc Natl Acad Sci U S A. 1987 Apr;84(7):2024–2028. doi: 10.1073/pnas.84.7.2024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pfeilschifter J., Seyedin S. M., Mundy G. R. Transforming growth factor beta inhibits bone resorption in fetal rat long bone cultures. J Clin Invest. 1988 Aug;82(2):680–685. doi: 10.1172/JCI113647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Poli V., Balena R., Fattori E., Markatos A., Yamamoto M., Tanaka H., Ciliberto G., Rodan G. A., Costantini F. Interleukin-6 deficient mice are protected from bone loss caused by estrogen depletion. EMBO J. 1994 Mar 1;13(5):1189–1196. doi: 10.1002/j.1460-2075.1994.tb06368.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Robey P. G., Young M. F., Flanders K. C., Roche N. S., Kondaiah P., Reddi A. H., Termine J. D., Sporn M. B., Roberts A. B. Osteoblasts synthesize and respond to transforming growth factor-type beta (TGF-beta) in vitro. J Cell Biol. 1987 Jul;105(1):457–463. doi: 10.1083/jcb.105.1.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rodan G. A., Martin T. J. Role of osteoblasts in hormonal control of bone resorption--a hypothesis. Calcif Tissue Int. 1981;33(4):349–351. doi: 10.1007/BF02409454. [DOI] [PubMed] [Google Scholar]
  40. Rosen D., Miller S. C., DeLeon E., Thompson A. Y., Bentz H., Mathews M., Adams S. Systemic administration of recombinant transforming growth factor beta 2 (rTGF-beta 2) stimulates parameters of cancellous bone formation in juvenile and adult rats. Bone. 1994 May-Jun;15(3):355–359. doi: 10.1016/8756-3282(94)90300-x. [DOI] [PubMed] [Google Scholar]
  41. Sandberg M., Vuorio T., Hirvonen H., Alitalo K., Vuorio E. Enhanced expression of TGF-beta and c-fos mRNAs in the growth plates of developing human long bones. Development. 1988 Mar;102(3):461–470. doi: 10.1242/dev.102.3.461. [DOI] [PubMed] [Google Scholar]
  42. Schneider H. G., Michelangeli V. P., Frampton R. J., Grogan J. L., Ikeda K., Martin T. J., Findlay D. M. Transforming growth factor-beta modulates receptor binding of calciotropic hormones and G protein-mediated adenylate cyclase responses in osteoblast-like cells. Endocrinology. 1992 Sep;131(3):1383–1389. doi: 10.1210/endo.131.3.1324161. [DOI] [PubMed] [Google Scholar]
  43. Seitz P. K., Zhu B. T., Cooper C. W. Effect of transforming growth factor beta on parathyroid hormone receptor binding and cAMP formation in rat osteosarcoma cells. J Bone Miner Res. 1992 May;7(5):541–546. doi: 10.1002/jbmr.5650070510. [DOI] [PubMed] [Google Scholar]
  44. Seyedin S. M., Thomas T. C., Thompson A. Y., Rosen D. M., Piez K. A. Purification and characterization of two cartilage-inducing factors from bovine demineralized bone. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2267–2271. doi: 10.1073/pnas.82.8.2267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Sillence D. O., Ritchie H. E., Selby P. B. Animal model: skeletal anomalies in mice with cleidocranial dysplasia. Am J Med Genet. 1987 May;27(1):75–85. doi: 10.1002/ajmg.1320270109. [DOI] [PubMed] [Google Scholar]
  46. Tashjian A. H., Jr, Voelkel E. F., Lazzaro M., Singer F. R., Roberts A. B., Derynck R., Winkler M. E., Levine L. Alpha and beta human transforming growth factors stimulate prostaglandin production and bone resorption in cultured mouse calvaria. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4535–4538. doi: 10.1073/pnas.82.13.4535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tran S., Hall B. K. Growth of the clavicle and development of clavicular secondary cartilage in the embryonic mouse. Acta Anat (Basel) 1989;135(3):200–207. doi: 10.1159/000146754. [DOI] [PubMed] [Google Scholar]
  48. Vermeulen A. H., Vermeer C., Bosman F. T. Histochemical detection of osteocalcin in normal and pathological human bone. J Histochem Cytochem. 1989 Oct;37(10):1503–1508. doi: 10.1177/37.10.2789247. [DOI] [PubMed] [Google Scholar]
  49. Weibel E. R. Stereological principles for morphometry in electron microscopic cytology. Int Rev Cytol. 1969;26:235–302. doi: 10.1016/s0074-7696(08)61637-x. [DOI] [PubMed] [Google Scholar]
  50. Whyte M. P., Bergfeld M. A., Murphy W. A., Avioli L. V., Teitelbaum S. L. Postmenopausal osteoporosis. A heterogeneous disorder as assessed by histomorphometric analysis of Iliac crest bone from untreated patients. Am J Med. 1982 Feb;72(2):193–202. doi: 10.1016/0002-9343(82)90810-5. [DOI] [PubMed] [Google Scholar]
  51. Wronski T. J., Lowry P. L., Walsh C. C., Ignaszewski L. A. Skeletal alterations in ovariectomized rats. Calcif Tissue Int. 1985 May;37(3):324–328. doi: 10.1007/BF02554882. [DOI] [PubMed] [Google Scholar]
  52. Wronski T. J., Walsh C. C., Ignaszewski L. A. Histologic evidence for osteopenia and increased bone turnover in ovariectomized rats. Bone. 1986;7(2):119–123. doi: 10.1016/8756-3282(86)90683-6. [DOI] [PubMed] [Google Scholar]
  53. Yee J. A., Yan L., Dominguez J. C., Allan E. H., Martin T. J. Plasminogen-dependent activation of latent transforming growth factor beta (TGF beta) by growing cultures of osteoblast-like cells. J Cell Physiol. 1993 Dec;157(3):528–534. doi: 10.1002/jcp.1041570312. [DOI] [PubMed] [Google Scholar]
  54. de Martin R., Haendler B., Hofer-Warbinek R., Gaugitsch H., Wrann M., Schlüsener H., Seifert J. M., Bodmer S., Fontana A., Hofer E. Complementary DNA for human glioblastoma-derived T cell suppressor factor, a novel member of the transforming growth factor-beta gene family. EMBO J. 1987 Dec 1;6(12):3673–3677. doi: 10.1002/j.1460-2075.1987.tb02700.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES