Summary
Normal transverse growth of long bones is by periosteal appositional bone formation, balanced by endosteal resorption. Changes in the distribution of cells that are expressing collagen mRNAs during growth were determined using digoxigeninlabelled riboprobes. In neonatal rabbit tibiae osteoblasts expressing type I collagen mRNA are found on periosteal, and at early stages on endosteal, bone surfaces and lining peripheral cavities. Occasional osteocytes express type I collagen mRNA very weakly. The pattern is disrupted when transforming growth factor-β2 (TGF-β2) is injected daily into the periosteum of neonatal animals; there is increased bone, and later cartilage, formation. Three injections of 20 ng TGF-β2 onto the tibia of 3-day-old rabbits led to an increase of periosteal osteoblasts that express the mRNA for type I collagen. Some endosteal osteoblasts and osteocytes in newly-formed peripheral woven bone also express the mRNA. After five injections chondrocytes expressing type II collagen mRNA are found around the injection site. Similar injections of TGF-β2 in old rabbits induce only fibrous tissue within which some cells express type I collagen mRNA. This precise localization of mRNAs shows that the expression of type I or II collagen mRNA is here restricted to osteoblasts and chondrocytes, respectively.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Benya, P. D. & Brown, P. D. (1986) Modulation of the chondrocyte phenotype in vitro. In Articular Cartilage Biochemistry (edited by) Keuttner, K. E., Schleyerbach, R. & Hascall, V. C.), pp. 219–30. New York: Raven Press.
Ber, R., Kubota, T., Sodek, J. & Aubin, J. E. (1991) Effects of transforming growth factor-beta on normal clonal bone cell populations. Biochem. Cell Biol. 9, 132–40.
Bland, Y. S., Critchlow, M. A. & Ashhurst, D. E. (1991) Digoxigenin as a probe label for in situ hybridization on skeletal tissues. Histochem. J. 23, 415–8.
Castagnola, P., Dozin, B., Moro, G. & Cancedda, R. (1988) Changes in the expression of collagen genes show two stages in chondrocyte differentiation in vitro. J. Cell Biol. 106, 461–7.
Critchlow, M. A., Bland, Y. S. & Ashhurst, D. E. (1994) The effects of age on the response of rabbit periosteal osteoprogenitor cells to exogenous transforming growth factor-β2. J. Cell Sci. 107, 499–516.
Futura, Y., Shinohara, T., Sano, K., Meguro, M. & Nagashima, K. (1990) In situ hybridization with digoxigenin-labelled DNA probes for detection of viral genomes. J. Clin. Pathol. 43, 806–9.
Joyce, M. E., Roberts, A. B., Sporn, M. B. & Bolander, M. E. (1990) Transforming growth factor-β and the initiation of chondrogenesis and osteogenesis in the rat femur. J. Cell Biol. 110, 2195–207.
Mackie, E. J. & Trechsel, U. (1990) Stimulation of bone formation in vivo by transforming growth factor-β: remodeling of woven bone and lack of inhibition by indomethacin. Bone 11, 295–300.
Mäkelä, J. R., Raassina, M., Virta, A. & Vuorio, E. (1988) Human proα1(I) collagen: cDNA sequence for the C-propeptide domain. Nucleic Acids Res. 16, 349.
Marcelli, C., Yates, A. J. & Mundy, G. R. (1990) In vivo effects of human recombinant transforming growth factor β on bone turnover in normal mice. J. Bone Min. Res. 5, 1087–96.
Martinez-Montero, J. C., Herrington, C. S., Stickland, J., Sawyer, H., Evans, M., Flannery, D. M. J. & Mcgee, J. O'D. (1991) Model systems for optimising mRNA non-isotopic in situ hybridization: riboprobe detection of lysozyme mRNA in archival gut biopsy specimens. J. Clin. Pathol. 44, 835–9.
Morris, R. G., Arends, M. J., Bishop, P. E., Sizer, K., Duvalle, E. & Bird, C. C. (1990) Sensitivity of digoxigenin and biotin labelled probes for detection of human papillomavirus by in situ hybridization. J. Clin. Pathol. 43, 800–5.
Mundlos, S., Engel, H., Michel-Behnke, I. & Zabel, B. (1990) Distribution of type I and type II collagen gene expression during the development of human long bones. Bone 11, 275–9.
Noda, M. & Camilliere, J. J. (1989) In vivo stimulation of bone formation by transforming growth factor-β. Endocrinology 124, 2991–4.
Owen, M. (1963) Cell population kinetics of an osteogenic tissue — I. J. Cell Biol. 19, 19–32.
Owen, M. & Macpherson, S. (1963) Cell population kinetics of an osteogenic tissue — II. J. Cell Biol. 19, 33–44.
Page, M., Hogg, J. & Ashhurst, D. E. (1986) The effects of mechanical stability on the macromolecules of the connective tissue matrices produced during fracture healing. I. The collagens. Histochem. J. 18, 251–65.
Sandberg, M. & Vuorio, E. (1987) Localization of types I, II and III collagen mRNAs in developing human skeletal tissues by in situ hybridization. J. Cell Biol. 104, 1077–84.
Sandberg, M., Autio-Harmainen, H. & Vuorio, E. (1988) Localization of the expression of types I, III and IV collagen, TGF-β1 and c-fos genes in developing human calvarial bones. Develop. Biol. 130, 324–34.
Sandell, L. J., Sugai, J. V. & Trippel, S. B. (1994) Expression of collagens I, II, X and XI and aggrecan mRNAs by bovine growth plate chondrocytes in situ. J. Orthop. Res. 12, 1–14.
Shroff, B., Smith, T., Norris, K., Pileggi, R. & Sauk, J. J. (1993) HSP 47 is localized to regions of type I collagen production in developing murine femurs and molars. Conn. Tissue Res. 29, 273–86.
Strong, D. D., Beachler, A. L., Wergedal, J. E. & Linkhart, T. A. (1991) Insulin like growth factor II and transforming growth factor β regulate collagen expression in human osteoblast like cells in vitro. J. Bone Min. Res. 6, 15–23.
Tanaka, T., Taniguchi, Y., Gotoh, K., Satoh, R., Inazu, M. & Ozawa, H. (1993) Morphological study of recombinant human transforming growth factor-β1-induced intramembranous ossification in neonatal rat parietal bone. Bone 14, 117–23.
Tonna, E. A. (1961) The cellular complement of the skeletal system studied autoradiographically with tritiated thymidine (H3TDR) during growth and aging. J. Biophys. Biochem. Cytol. 9, 813–24.
Tonna, E. A. & Cronkite, E. P. (1962) An autoradiographic study of periosteal cell proliferation with tritiated thymidine. Lab. Invest. 11, 455–62.
Young, R. W. (1962) Regional differences in cell generation time in growing rat tibiae. Exp. Cell Res. 26, 562–7.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Critchlow, M.A., Bland, Y.S. & Ashhurst, D.E. The expression of collagen mRNAs in normally developing neonatal rabbit long bones and after treatment of neonatal and adult rabbit tibiae with transforming growth factor-β2. Histochem J 27, 505–515 (1995). https://doi.org/10.1007/BF00174323
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00174323