Analysis of Embryonic Cartilage and Bone Induction in a Defined Culture System

  • Harold C. Slavkin
  • Malcolm L. Snead
  • Wen Luo
  • Pablo BringasJr.
  • Shigeshi Kikunaga
  • Yasuyuki Sasano
  • Conny Bessem
  • Mark Mayo
  • Mary MacDougall
  • Leslie B. Rall
  • Daniel Rappolee
  • Zena Werb
Part of the NATO ASI Series book series (NSSA, volume 184)


The primary means by which vertebrate mineralized tissues become determined during development is by interaction between different regions of the embryo, a process known as embryonic induction. Numerous examples of embryonic induction were intensively studied problems in developmental biology during the first 70 years of the 20th century (see reviews by Spemann, 1938, Grobstein, 1967; Hall, 1988). Progress in the last few years has in part been the result of applications of recombinant DNA technology to classical questions in the field of embryonic induction (e.g. see recent reviews by Gurdon, 1987; 1988; Edelman, 1988). The key issues appear to be when, where and which sequence of regulatory factor expression activate signal transduction processes resulting in the allocation, determination and differentiation of specific phenotypes. There are probably multiple signals and multiple receptors required for inductive processes.


Branchial Arch Polypeptide Growth Factor Bone Induction Mesoderm Induction Embryonic Induction 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anderson, H.C., 1976, Osteogenic epithelial-mesenchymal cell interactions, Clin. Orthop. Rel. Res., 119: 211.Google Scholar
  2. Beck, F., Samani, N.J., Penschow, J.D., Thorley, B., Tregear, G.W., and Coghlan, J.P., 1987, Histochemical localization of IGF-I and -II mRNA in the developing rat embryo, Development, 101: 175.PubMedGoogle Scholar
  3. Brachmann, R., Lindquist, P.B., Nagashima, M., Kohr, W., Lipari, T., Napier, M., and Derynck, R., 1989, Transmembrane TGF-alpha precursors activate EGF/TGFalpha receptors, Cell, 56: 691.Google Scholar
  4. Canalis, E., 1985, Effect of growth factors on bone cell replication and dfferentiation, Clin. Orthop. Relat. Res., 193: 246.PubMedGoogle Scholar
  5. Caplan, A.I. and Pechak, D.G., 1987, The cellular and molecular embryology of bone formation, in: “Bone And Mineral Research/5,” W.A. Peck, ed., Elsevier, Amsterdam.Google Scholar
  6. Centrella, M. and Canalis, E., 1985, Transforming and nontransforming growth factors are present in medium conditioned by fetal rat calvariae, Proc. Natl. Acad. Sci., USA, 82: 7335.CrossRefGoogle Scholar
  7. Dale, L. and Slack, J.M.V., 1987, Regional specification within the mesoderm of early embryos of Xenopus laevis, Development, 100: 279.PubMedGoogle Scholar
  8. Derynck, R., 1988, Transforming growth factor alpha, Cell, 54: 593.PubMedCrossRefGoogle Scholar
  9. Edelman, G.M., 1988, “Topobiology”, Basic Books, Inc., New York.Google Scholar
  10. Gans, C., 1988, Craniofacial growth, evolutionary questions, Development, 103: 3.PubMedGoogle Scholar
  11. Greenwald, I., 1985, Lin-2, a nematode homeotic gene is homologous to a set of mammalian proteins that includes epidermal growth factor, Cell, 43: 583.PubMedCrossRefGoogle Scholar
  12. Grobstein, C., 1967, Mechanisms of organogenetic tissue interaction, Natl. Cancer Inst. Monogr., 26: 279.PubMedGoogle Scholar
  13. Gurdon, J.B., 1987, Embryonic induction-molecular prospects, Development, 99:285. Gurdon, J.B., 1988, A community effect in animal development, Nature, 336:772. Hall, B.K., 1980, Tissue interactions and the initiation of osteogenesis and chondrogenesis in the neural crest-derived mandibular skeleton of the embryonic mouse as seen in isolated murine tissues and in recombination of murine and avian tissues, J. Embryol. Exp. Morph., 58: 251.Google Scholar
  14. Hall, B.K., 1981, The induction of neural crest-derived cartilage and bone by embryonic epithelia: an analysis of the mode of action of an epithelialmesenchymal interaction, J. Embryol. Exp. Morph., 64: 305.PubMedGoogle Scholar
  15. Hall, B.K., 1982, The role of tissue interactions in the growth of bone. in: “Factors and Mechanisms Influencing Bone Growth,” A.S. Dixon and B.G. Sarnat, eds., Alan R. Liss, Inc., New York.Google Scholar
  16. Hall, B.K., 1984, Genetic and epigenetic control of connective tissues in the cranial structures. Birth Defects: Original Article Series, 20 (3): 1.Google Scholar
  17. Hall, B.K., 1987, Initiation of chondrogenesis from somitic, limb and craniofacial mesenchyme: search for a common mechanism, in: “Somites in Developing Embryos,” R. Bellairs, D.A. Ede and J.W. Lash, eds., Plenum, New York.Google Scholar
  18. Hall, B.K., I988a, Patterning of connective tissues in the head: discussion report, Development, 103: 171.Google Scholar
  19. Hall, B.K., 1988b, The embryonic development of bone, American. Sci., 76:174. Hall, B.K., Van Exan, R., and Brunt, S., 1983, Retention of epithelial basal laminaGoogle Scholar
  20. allows isolated mandibular mesenchyme to form bone, J. Craniofac. Genet. & Develop. Biol.,3:253.Google Scholar
  21. Han, V.K.M., D’Ercole, J., and Lund, P.K., 1987, Cellular localization of somatomedin (insulin-like growth factor)messenger RNA in the human fetus, Science, 236: 193.PubMedCrossRefGoogle Scholar
  22. Hauschka, P.V., Mavrakos, A.E., Iafrati, M.D., Doleman, S.E., and Klagsbrun, M., 1986, Growth factors in bone matrix. Isolation of multiple types by affinity chromatography on heparin-Sepharose, J. Biol. Chem., 261: 1 2665.Google Scholar
  23. Hauschka, P.V., Chen, T.L., and Mavrakos, A.E., 1988, Polypeptide growth factors in bone matrix, in: “Cell and Molecular Biology of Vertebrate Hard Tissues,” G.A. Rodan, ed., Wiley, Chichester.Google Scholar
  24. Jacobsen, W. and Fell, H.B., 1941, The developmental mechanics and potencies of theGoogle Scholar
  25. undifferentiated mesenchyme of the mandible, Quart. J. Micro. Sci.,82:563.Google Scholar
  26. Jones, F.S., Burgoon, M.P., Hoffman, S., Crossin, K.L., Cunningham, B.A., and Edelman, G.M., 1988, A cDNA clone for cytotactin contains sequences similar to epidermal growth factor-like repeats and segments of fibronectin and fibrinogen, Proc. Natl. Acad. Sci., USA, 85: 2186.PubMedCrossRefGoogle Scholar
  27. Jones, F.S., Hoffman, S., Cunningham, B.A., and Edelman, G.M., 1989, A detailed structural model of cytotactin: protein homologies, alternative RNA splicing and binding regions, Proc. Natl. Acad. Sci. USA, 86: 1905.PubMedCrossRefGoogle Scholar
  28. Kawamura, M. and Urist, M.R., 1988, Growth factors, mitogens, cytokines and bone morphogenetic protein in induced chondrogenesis in tissue culture, Develop. Biol., 130: 435.PubMedCrossRefGoogle Scholar
  29. Kimelman, D. and Kirschner, M., 1987, Synergistic induction of mesoderm by FGF and TGF-beta and the identification of an mRNA coding for FGF in the early Xenopus embryo, Cell, 51: 869.PubMedCrossRefGoogle Scholar
  30. Lian, J., Stewart, C., Puchacz, E., Mackowiak, S., Shalhoub, V., Collart, D., Zambetti, G., and Stein, G., 1989, Structure of the rat osteocalcin gene and regulation of vitamin D-dependent expression, Proc. Natl. Acad. Sci, USA, 86: 1143.PubMedCrossRefGoogle Scholar
  31. Lumsden, A.G.S., 1988, Spatial organization of the epithelium and the role of neural crest cells in the initiation of the mammalian tooth germ, Development, 103: 155.PubMedGoogle Scholar
  32. Mackie, E.J., Thesleff, I., and Chiquet-Ehrismann, R., 1987, Tenascin is associated with chondrogenic and osteogenic differentiation in vivo and promotes chondrogenesis in vitro, J. Cell Biol., 105: 2569.PubMedCrossRefGoogle Scholar
  33. Mohan, S., Linkhart, T.A., Jennings, J.C., and Baylink, D.J., 1987, Identification and quantitation of four distinct growth factors stored in human bone matrix, J. Bone Miner. Res., 2(Suppl. 1):44. •Google Scholar
  34. Nakano, T., Kimoto, S., Tanikawa, K., Kim, K.T., Higaki, M., Kawase, T., and Saito, S., 1989, Identification of osteoblast-specific monoclonal antibodies, Calcif. Tiss Int., 44: 220.CrossRefGoogle Scholar
  35. Nilsson, A., Isgaard, J., Lindahl, A., Dahlstrom, A., Skottner, A., Isaksson, O.G.P., 1986, Regulation by growth hormone of number of chondrocytes containing IGF-I in rat growth plate, Science, 233: 571–574.PubMedCrossRefGoogle Scholar
  36. Otte, A.P., Koster, C.H., Snoek, G.T., and Durston, A.J., 1988, Protein kinase C mediates neural induction in Xenopus laevis, Nature, 334: 618.PubMedCrossRefGoogle Scholar
  37. Panayotou, G., End, P., Aumailley, M., Timpl, R., and Engel, J., 1989, Domains of laminin with growth-factor activity, Cell, 56: 93.PubMedCrossRefGoogle Scholar
  38. Pearson, C.A., Pearson, D., Shibahara, S., Hofsteenge, J., and Chiquet-Ehrismann, R., 1988, Tenascin: cDNA cloning and induction by TGF-beta, EMBO J., 7:2977. Pechak, D.G., Kujawa, M.J., and Caplan, A.I., 1986, Morphology of bone development and bone remodeling in embryonic chick limbs, Bone, 7: 459.Google Scholar
  39. Pedersen, R.A., 1988, Early mammalian embryogenesis, in: “The Physiology of Reproduction,” E. Knobil and J. Neill et al., eds., Raven Press, Ltd., New York.Google Scholar
  40. Pfeilschifter, J., D’Souza, S., and Mundy, G.R., 1986, Transforming growth factor beta is released from resorbing bone and stimulates osteoblast activity, J. Bone Miner. Res., 1 (Suppl.): 294.Google Scholar
  41. Rappolee, D.A., Mark, D., Banda, M.J., and Werb, Z., 1988a, Wound macrophages express TGF-alpha and other growth factors in vivo: analysis by mRNA phenotyping, Science, 241: 708.PubMedCrossRefGoogle Scholar
  42. Rappolee, D.A., Brenner, C.A., Schultz, R., Mark, D., and Werb, Z., 1988b, Developmental expression of PDGF, TGF-alpha and TGF-beta genes in preimplantation mouse embryos, Science, 241: 1823.PubMedCrossRefGoogle Scholar
  43. Rappolee, D.A., Wang, A., Mark, D., and Werb, Z., 1989, Novel method for studying mRNA phenotypes in single or small numbers of cells, J. Cell. Biochem., 39: 1.PubMedCrossRefGoogle Scholar
  44. Raisz, L.G., 1988, Hormonal regulation of bone growth and remodelling, in: “Cell and Molecular Biology of Vertebrate Hard Tissues,” G.A. Rodan, ed., Wiley, Chichester.Google Scholar
  45. Rall, L.B., Scott, J., Bell, G.I., Crawford, R.J., Penschow, J.D., Niall, H.D., and Coghlan, J.P., 1985, Mouse prepro-epidermal growth factor synthesis by the kidney and other tissues, Nature, 313: 228PubMedCrossRefGoogle Scholar
  46. Robey, P.G., Young, M.F., Flanders, K.C., Roche, N.S., Kondaiah, P., Reddi, A.H., Termine, J.D., Sporn, M.B., and Roberts, A.B., 1987, Osteoblasts synthesize and respond to transforming growth factor-type beta (TGF-beta) in vitro, J. Cell Biol., 105: 457.PubMedCrossRefGoogle Scholar
  47. Rodan, G.A., Heath, J.K., Yoon, K., Noda, M., and Rodan, S.B., 1988, Diversity of the osteoblastic phenotype, in: “Cell and Molecular Biology of Vertebrate Hard Tissues,” G.A. Rodan, ed., Wiley, Chichester.Google Scholar
  48. Rutter, W.J., and Pictet, R.L., 1976, Hormone-like factor(s) in mesenchymal-epithelial interactions during embryonic development, in: “Embryogenesis in Mammals,” K.Elliot and M. O’Connor, eds., Wiley, New York.Google Scholar
  49. Sandberg, M., Vuorio, T., Hirvonen, H., Alitalo, K., and Vuorio, E., 1988, Enhanced expression of TGF-beta and c-fos mRNAs in the growth plates of developing human long bones, Development, 102: 461–470.Google Scholar
  50. Sandy, J.R., Meghji, S., Scutt, A.M., Harvey, W., Harris, M., and Meikle, M.C., 1989, Murine osteoblasts release bone-resorbing factors of high and low molecular weights: stimulation by mechanical deformation, Bone and Mineral, 5: 155.PubMedCrossRefGoogle Scholar
  51. Scott, J., Urdea, M., Quiroga, M., Sanchez-Pescador, R., Fong, N., Selby, M., Rutter, W.J., and Bell, G.í.,1983, Structure of a mouse submaxillary messenger RNA encoding epidermal growth factor and seven related proteins, Science, 221: 236.Google Scholar
  52. Seyedin, S.M., Segarini, P.R., Rosen, D.M., Thompson, A.Y., Bentz, H., and Graycar, J., 1987, Cartilage-inducing factor-B is a unique protein structurally and functionally related to transforming growth factor-beta, J. Biol. Chem., 262: 1946.PubMedGoogle Scholar
  53. Shapiro, I.M., Golub, E.E., Chance, B., Piddington, C., Oshima, O., Tuncay, O.C., and Haselgrove, J.C., 1988, Linkage between energy status of perivascular cells and mineralization of the chick growth cartilage, Develop. Biol., 129: 372.PubMedCrossRefGoogle Scholar
  54. Sharpe, C.R., Fritz, A., De Robertis, E.M., and Gurdon, J.B., 1987, A homeoboxcontaining marker of posterior neural differentiation shows the importance of predetermination in neural induction, Cell, 50: 749.Google Scholar
  55. Slack, J.M.W., Darlington, B.G., Heath, J.K., and Godsave, S.F., 1987, Mesoderm induction in early Xenopus embryos by heparin-binding growth factors, Nature, 326: 197.PubMedCrossRefGoogle Scholar
  56. Slavkin, H.C., 1978, Mandibular morphogenesis, in: “Reconstruction of Jaw Deformities,” L.A. Whitaker, ed., C.V. Mosby Co., St. Louis.Google Scholar
  57. Slavkin, H.C., Bringas, P., Cummings, E.C., and Grodin, M.S., 1982a, Murine mandibular chondrogenesis and osteogenesis in a serumless, chemically-defined medium, in: “Chemistry and Biology of Mineralized Connective Tissues,” A. Veis, ed., Elsevier/North holland, New York.Google Scholar
  58. Slavkin, H.C., Honig, L.S., and Bringas, P. 1982b, Experimental dissection of avian and murine tissue interactions using organ culture in a serumless medium free from exogenous (nondefined) factors, in:“Factors and Mechanisms Influencing Bone Growth,” A.D. Dixon and B.G. Sarnat, eds., Alan R. Liss, Inc., New York.Google Scholar
  59. Slavkin, H.C., Bringas, P., Sasano, Y., and Mayo, M., (in press) Early embryonic mouse mandibular morphogenesis and cytodifferentiation in serumless, chemically-defined medium: A model for studies of autocrine and/or paracrine regulatory factors, J. Craniofac. Genet. & Develop. Biol. Google Scholar
  60. Slavkin, H.C., Sasano, Y., Kikunaga, S., Bessern, C., Bringas, P., Mayo, M., Luo, W., Mak, G., Rall, L., and Snead, M.L. (in press) Cartilage, bone and tooth induction during early embryonic mouse mandibular morphogenesis using serumless, chemically-defined medium, Conn. Tiss. Res. Google Scholar
  61. Smith, J.C., 1987, A mesoderm-inducing factor is produced by a Xenopus cell line, Development, 99: 3.PubMedGoogle Scholar
  62. Smith, L. and Thorogood, P., 1983, Transfilter studies on the mechanism of epitheliomesenchymal interaction leading to chondrogenic differentiation of neural crest cells, J. Embryol. Exp. Morph., 75: 165.Google Scholar
  63. Smith, J.C., Yagoob, M., and Symes, K., 1988, Purification, partial characterization and biological effects of the XTC mesoderm-inducing factor, Development, 103: 591.PubMedGoogle Scholar
  64. Snead, M.L., Luo, W., Oliver, P. Nakamura, M., Don-Wheeler, G., Bessern, C., Bell, G.I., Rall, L.B., and Slavkin, H.C., (in press) Localization of epidermal growth factor precursor in tooth and lung during embryonic mouse development, Develop. Biol.Google Scholar
  65. Spemann, H, 1938, “Embryonic Induction and Development, ” Yale University Press, New Haven.Google Scholar
  66. Stylianopoulou, F., Efstratiadis, A., Herbert, J., and Pintar, J., 1988, Pattern of the insulin-like growth factor II gene expression during rat embryogenesis, Development, 103: 497–506.PubMedGoogle Scholar
  67. Termine, J.D., 1988, Non-collagen proteins in bone, in: “Cell and Molecular Biology of Vertebrate Hard Tissues,” G.A. Rodan, ed., Wiley, Chichester.Google Scholar
  68. Thorogood, P. 1988, The developmental specification of the vertebrate skull, Development,103:141.Google Scholar
  69. Tomkins, G.M., 1975, The metabolic code, Science, 189: 760.PubMedCrossRefGoogle Scholar
  70. Tyler, M.S. and Hall, B.K., 1977, Epithelial influences on skeletogenesis in the mandible of the embryonic chick, Anat. Rec., 188: 229.PubMedCrossRefGoogle Scholar
  71. Urist, M.L., DeLange, R.J., and Finerman, G.A.M., 1983, Bone cell differentiation and growth factors, Science, 220: 680.PubMedCrossRefGoogle Scholar
  72. Wahl, M.I., Nishibe, S., Suh, P.G., Rhee, S.G., and Carpenter, G., 1989, Epidermal growth factor stimulates tyrosine phosphorylation of phopholipase C-II independently of receptor internalization and extracellular calcium, Proc. Natl. Acad. Sci. USA, 86: 1568.PubMedCrossRefGoogle Scholar
  73. Weeks, D.L. and Melton, D.A., 1987, A maternal mRNA localized to the vegetal hemisphere in Xenopus eggs codes for a growth factor related to TGF-beta, Cell, 51: 861.PubMedCrossRefGoogle Scholar
  74. Wharton, K.A., Johansen, K.M., Xu, T., and Artavanis, T., 1985, Nucleotide sequence from the neurogenic locus Notch implies a gene product that shares homology with proteins containing EGF-like repeats, Cell, 43: 567.Google Scholar
  75. Wilcox, J.N. and Derynck, R., 1988, Developmental expression of transforming growth factors alpha and beta in mouse fetus, Molecular & Cellular Biology, 8: 3415.Google Scholar
  76. Wong, S.T., Winchell, L.F., McCune, B.K., Earp, H.S., Teixido, J., Massague, J., Herman, B., and Lee, D.C., 1989, The TGF-alpha precursor expressed on the cell surface binds to the EGF receptor on adjacent cells, leading to signal transduction Cell56:495. Google Scholar
  77. Wozney, J.M., Rosen, V., Celeste, A.J., Mitsock, L.M., Whitters, M.J., Kriz, R.W., Hewick, R.M., and Wang, E.A., 1988, Novel regulators of bone formation: molecular clones and activities, Science, 243: 1528CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1990

Authors and Affiliations

  • Harold C. Slavkin
    • 1
  • Malcolm L. Snead
    • 1
  • Wen Luo
    • 1
  • Pablo BringasJr.
  • Shigeshi Kikunaga
    • 1
  • Yasuyuki Sasano
    • 1
  • Conny Bessem
    • 1
  • Mark Mayo
    • 1
  • Mary MacDougall
    • 1
  • Leslie B. Rall
    • 2
  • Daniel Rappolee
    • 2
  • Zena Werb
    • 2
  1. 1.Laboratory for Developmental Biology Department of Basic Sciences School of DentistryUniversity of Southern CaliforniaLos AngelesUSA
  2. 2.Laboratory of Radiobiology & Environmental Health School of MedicineUniversity of California-San FranciscoSan FranciscoUSA

Personalised recommendations