This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Heikinheimo, K. Articles by Ruch, J.V. Search for Related Content PubMed PubMed Citation Articles by Heikinheimo, K. Articles by Ruch, J.V. Social Bookmarking                         What's this?

The Activin-binding Protein Follistatin is Expressed in Developing Murine Molar and Induces Odontoblast-like Cell Differentiation in vitro

Institut de Biologie Medicale, Université Louis Pasteur, Faculte de Medecine, INSERM U 424, Strasbourg, France, Institute of Dentistry, University of Turku, FIN-20520 Turku, Finland

C. Begue-Kirn

Institut de Biologie Medicale, Université Louis Pasteur, Faculte de Medecine, INSERM U 424, Strasbourg, France

O. Ritvos

University of Helsinki, Haartman Institute, Department of Bacteriology and Immunology, FIN-00014 Helsinki, Finland

T. Tuuri

University of Helsinki, Haartman Institute, Department of Bacteriology and Immunology, FIN-00014 Helsinki, Finland

J.V. Ruch

Institut de Biologie Medicale, Université Louis Pasteur, Faculte de Medecine, INSERM U 424, Strasbourg, France

It has recently been shown that mice deficient in activin-βA subunits and follistatin exhibit major defects in dentition. To increase understanding of the roles played by these molecules during tooth development, we determined the temporospatial expression of activin-βA subunit and follistatin messenger RNA and their corresponding proteins in developing murine molars (between day E 14 and 2 days after birth). The effects of recombinant human activin A and its binding protein follistatin on odontoblast differentiation were also studied in cultures of dental papillae (DP) isolated from the mandibular first molars of E-17-day mice. In situ hybridization indicated that transcripts for activin-βA subunit were abundant in pre-odontoblasts at the tips of forming cusps prior to odontoblast terminal differentiation, and transcripts for follistatin in overlying inner enamel epithelial cells (pre-ameloblasts). Pre-odontoblasts were also weakly immunoreactive in relation to activin-βA subunit, pre-ameloblasts in relation to follistatin. When follistatin was added at different concentrations to a DP culture model (2-14 nmol/DP) together with heparin at constant concentration, differentiation of odontoblast-like cells was induced, as evidenced by polarization and deposition of extracellular matrix in vitro, to extents depending on the follistatin concentration. In contrast, the addition of activin A (2 nmol/DP) had no effect on the differentiation parameters studied. These findings suggest that the activin-follistatin system regulates odontoblast differentiation during tooth development. In particular, we suggest that binding of endogenous activin A by follistatin may allow odontoblast terminal differentiation to occur.

Key Words: activin A • follistatin • immunohistochemistry • in situ hybridization • odontoblasts • dental papilla culture.

REFERENCES

• Albano RM, Groome N., Smith JC (1993). Activins are expressed in preimplantation mouse embryos and in ES and EC cells and are regulated on their differentiation. Development 117:711-723.[Abstract]
• Attisano L., Wrana JL, Montalvo E., Massague J. (1996). Activation of signalling by the activin receptor complex. Mol Cell Biol 16:1066-1073.[Abstract]
• Begue-Kirn C., Smith AJ, Ruch JV, Wozney JM, Purchio A., Hartmann D., et al. (1992). Effects of dentin proteins, transforming growth factor β1 (TGF-β1) and bone morphogenetic protein 2 (BMP2) on the differentiation of odontoblast in vitro. Int J Dev Biol 36:491-503.[Medline] [Order article via Infotrieve]
• Begue-Kirn C., Smith AJ, Loriot M., Kupferle C., Ruch JV, Lesot H. (1994). Comparative analysis of TGFβs, BMPs, IGFs, msxs, fibronectin, osteonectin and bone sialoprotein gene expressions during normal and in vitro induced odontoblast differentiation. Int j Dev Biol 38:405-420.[Medline] [Order article via Infotrieve]
• Chai Y., Mah A., Crohin C., Groff S., Bringas P. Jr, Le T., et al. (1994). Specific transforming growth factor-beta subtypes regulate embryonic mouse Meckel's cartilage and tooth development. Dev Biol 162:85-103.[CrossRef][Medline] [Order article via Infotrieve]
• Chen TL, Bates RL, Dudley A., Hammonds RG Jr, Amento EP (1991). Bone morphogenetic protein-2b stimulation of growth and osteogenic phenotypes in rat osteoblast-like cells: comparison with TGF-β1. J Bone Miner Res 6:1387-1393.[Medline] [Order article via Infotrieve]
• Decimo D., Georges-Labouesse E., Dolle P. (1995). In situ hybridization of nucleic acid probes to cellular RNA. In: Gene probes. A practical approach. Vol. 2. Hames BD, Higgins SJ, editors. Oxford, England: Oxford University Press, pp. 183-210.
• de Winter JP, ten Dijke P., de Vries CJ, van Achterberg TA, de Waele P., Huylebroeck D., et al. (1996). Follistatins neutralize activin bioactivity by inhibition of activin binding to its type II receptors. Mol Cell Endocrinol 116:105-114.[CrossRef][Medline] [Order article via Infotrieve]
• Feijen A., Goumans MJ, van den Eijnden-van Raaij AJ (1994). Expression of activin subunits, activin receptors and follistatin in postimplantation mouse embryos suggest specific development functions for different activins. Development 120:3621-3637.[Abstract]
• Groome N., Lawrence M. (1991). Preparation of monoclonal antibodies to the beta A subunit of ovarian inhibin using a synthetic peptide immunogen. Hybridoma 10:309-316.[Medline] [Order article via Infotrieve]
• Hashimoto M., Shoda A., Inoue S., Yamada R., Kondo T., Sakurai T., et al. (1992a). Functional regulation of osteoclastic cells by the interaction of activin-A with follistatin. J Biol Chem 267:4999-5004.[Abstract/Free Full Text]
• Hashimoto M., Nakamura T., Inoue S., Kondo T., Yamada R., Eto Y., et al. (1992b). Follistatin is a developmentally regulated cytokine in neural differentiation. J Biol Chem 267:7203-7206.[Abstract/Free Full Text]
• Heikinheimo K. (1994). Stage-specific expression of decapentaplegic-Vg-related genes 2, 4, and 6 (bone morphogenetic proteins 2, 4, and 6) during human tooth morphogenesis. J Dent Res 73:590-597.
• Heikinheimo K., Happonen R-P., Miettinen PJ, Ritvos O. (1993). Transforming growth factor beta 2 in epithelial differentiation of developing teeth and odontogenic tumors. J Clin Invest 91:1019-1027.[Medline] [Order article via Infotrieve]
• Inouye S., Guo Y., DePaolo L., Shimonaka M., Ling N., Shimasaki S. (1991). Recombinant expression of human follistatin with 315 and 288 amino acids: chemical and biological comparison with native porcine follistatin. Endocrinology 129:815-822.[Abstract/Free Full Text]
• Kingsley DM (1994). The TGF-β superfamily: new members, new receptors, and new genetic tests of function in different organisms. Gene Dev 8:133-146.[Free Full Text]
• Klein PS, Melton DA (1994). Hormonal regulation of embryogenesis: the formation of mesoderm in Xenopus laevis. Endocr Rev 15:326-341.[Abstract/Free Full Text]
• Mathews LS (1994). Activin receptors and cellular signalling by the receptor serine kinase family. Endocr Rev 15:310-325.[Abstract/Free Full Text]
• Matzuk MM, Kumar TR, Vassalli A., Bickenbach JR, Roop DR, Jaenisch R. (1995a). Functional analysis of activins during mammalian development. Nature 374:354-356.[CrossRef][Medline] [Order article via Infotrieve]
• Matzuk MM, Lu N., Vogel H., Sellheyer K., Roop DR, Bradley A. (1995b). Multiple defects and perinatal death in mice deficient in follistatin. Nature 374: 360-363.[CrossRef][Medline] [Order article via Infotrieve]
• Melton DA, Krieg PA, Rebagliati MR, Maniatis T., Zinn K., Green MR (1984). Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res 12:7035-7056.[Abstract/Free Full Text]
• Michel U., Farnsworth P., Findlay JK (1993). Follistatins: more than follicle-stimulating hormone suppressing proteins. Mol Cell Endocrinol 91:1-11.[CrossRef][Medline] [Order article via Infotrieve]
• Miettinen PJ, Heikinheimo K. (1992). Transforming growth factor-alpha (TGF-alpha) and insulin gene expression in human fetal pancreas. Development 114:833-840.[Abstract]
• Nakamura T., Takio K., Eto Y., Shibai H., Titani K., Sugino H. (1990). Activin-binding protein from rat ovary is follistatin. Science 247:836-838.[Abstract/Free Full Text]
• Nakamura T., Sugino K., Titani K., Sugino H. (1991). Follistatin, an activin-binding protein, associates with heparan sulphate chains of proteoglycans on follicular granulosa cells. J Biol Chem 266:19432-19437.[Abstract/Free Full Text]
• Nakashima M., Nagasawa H., Yamada Y., Reddi AH (1994). Regulatory role of transforming growth factor-β, bone morphogenetic protein-2 and protein-4 on gene expression of extracellular matrix proteins and differentiation of dental pulp cells. Dev Biol 162:18-28.[CrossRef][Medline] [Order article via Infotrieve]
• Nakatani A., Shimasaki S., DePaolo LV, Erickson GF, Ling N. ( 1991). Cyclic changes in follistatin messenger ribonucleic acid and its protein in the rat ovary during the estrous cycle. Endocrinol 129:603-611.[Abstract/Free Full Text]
• Paralkar VM, Nandedkar AK, Pointer RH, Kleinman HK, Reddi AH (1990). Interaction of osteogenin, a heparin binding bone morphogenetic protein, with type IV collagen. J Biol Chem 265:17281-17284.[Abstract/Free Full Text]
• Rentrop M., Knapp B., Winter H., Schweizer J. (1986). Aminoalkylsilane treated glass slides as support for in situ hybridization of keratin cDNAs to frozen tissue sections under varying fixation pretreatment conditions. Histochem J 18:271-276.[CrossRef][Medline] [Order article via Infotrieve]
• Ritvos O., Tuuri T., Eramaa M., Sainio K., Hilden K., Saxen L., et al. (1995). Activin disrupts epithelial branching morphogenesis in developing glandular organs of the mouse. Mech Dev 50:229-245.[CrossRef][Medline] [Order article via Infotrieve]
• Roberts VJ, Barth SL (1994). Expression of messenger ribonucleic acids encoding the inhibin/activin system during mid- and late-gestation rat embryogenesis. Endocrinol 134:914-923.[Abstract/Free Full Text]
• Roberts VJ, Sawchenko PE, Vale W. ( 1991). Expression of inhibin/activin subunit messenger ribonucleic acids during rat embryogenesis. Endocrinol 128:3122-3129.[Abstract/Free Full Text]
• Robertson DM, Klein R., de Vos FL, MacLachlan RI, Wettenhall RE, Hearn MT, et al. (1987). The isolation of polypeptides with FSH suppressing activity from bovine follicular fluid which are structurally different to inhibin. Biochem Biophys Res Commun 149:744-749.[CrossRef][Medline] [Order article via Infotrieve]
• Ruch JV, Karcher-Djuricic V., Thiebold J. (1976). Cell division and cytodifferentiation of odontoblasts. Differentiation 5:165-169.[CrossRef][Medline] [Order article via Infotrieve]
• Ruch JV, Lesot H., Begue-Kirn C. (1995). Odontoblast differentiation. Int J Dev Biol 39:51-68.[Medline] [Order article via Infotrieve]
• Schneyer AL, Rzucidlo DA, Sluss PM, Crowley WF Jr (1994). Characterization of unique binding kinetics of activin or inhibin in serum. Endocrinol 135:667-674.[Abstract]
• Schrewe H., Gendron-Maguire M., Harbison ML, Gridley T. (1994). Mice homozygous for a null mutation of activin βB are viable and fertile. Mech Dev 47:43-51.[CrossRef][Medline] [Order article via Infotrieve]
• Segarini PR (1993). TGF-β receptors, a complicated system of multiple binding proteins. Biochem Biophys Acta 1155:269-275.[Medline] [Order article via Infotrieve]
• Sugawara M., DePaolo L., Nakatani A., DiMarzo SJ, Ling N. ( 1990). Radioimmunoassay of follistatin: application for in vitro fertilization procedures. J Clin Endocrinol Metab 71:1672-1674.[Abstract/Free Full Text]
• Sumitomo S., Inouye S., Liu X.-J, Ling N., Shimasaki S. (1995). The heparin binding site of follistatin is involved in its interaction with activin. Biochem Biophys Res Commun 208:1-9.[CrossRef][Medline] [Order article via Infotrieve]
• Takuwa Y., Ohse C., Wang EA, Wozney JM, Yamashita K. (1991). Bone morphogenetic protein-2 stimulates alkaline phosphatase activity and collagen synthesis in cultured osteoblastic cells, MC3T3-E1. Biochem Biophys Res Commun 174:96-101.[CrossRef][Medline] [Order article via Infotrieve]
• ten Dijke P., Miyazono K., Heldin C.-H (1996). Signaling via hetero-oligomeric complexes of type I and type II serine/threonine kinase receptors. Cur Opin Cell Biol 8:139-145.[CrossRef][Medline] [Order article via Infotrieve]
• Thesleff I., Vaahtokari A., Partanen A-M (1995). Regulation of organogenesis. Common molecular mechanisms regulating the development of teeth and other organs. Int J Dev Biol 93:35-50.
• Thies RS, Bauduy M., Ashton BA, Kurtzberg L., Wozney JM, Rosen V. (1992). Recombinant human bone morphogenetic protein-2 induces osteoblast differentiation in W-20-17 stromal cells. Endocrinol 130:1318-1324.[Abstract/Free Full Text]
• Trowell OA (1954). A modified technique for organ culture in vitro. Exp Cell Res 46:246-248.[CrossRef]
• Tsuchida K., Vaughan JM, Wiater E., Gaddy-Kurten D., Vale WW ( 1995). Inactivation of activin-dependent transcription by kinase-deficient activin receptors. Endocrinol 136:5493-5503.[Abstract]
• Tuuri T., Eramaa M., Hilden K., Ritvos O. (1994). Activin-binding protein follistatin messenger ribonucleic acid secreted protein levels are induced by chorionic gonadotropin in cultured human granulosa-luteal cells. Endocrinol 135:2196-2203.[Abstract]
• Ueno N., Ling N., Ying SY, Esch F., Shimasaki S., Guillemin R. (1987). Isolation and partial characterization of follistatin: a single chain Mr 35,000 monomeric protein that inhibits the release of follicle-stimulating hormone. Proc Natl Acad Sci USA 80:8282-8286.
• Vaahtokari A., Aberg T., Jernvall J., Keranen S., Thesleff I. (1996). The enamel knot as a signaling center in the developing mouse tooth. Mech Dev 54:39-43.[CrossRef][Medline] [Order article via Infotrieve]
• Vainio S., Karavanova I., Jowett A., Thesleff I. (1993). Identification of BMP-4 as a signal mediating secondary induction between epithelial and mesenchymal tissues during tooth development. Cell 75:45-58.[CrossRef][Medline] [Order article via Infotrieve]
• Vale WA, Hsueh A., Rivier C., Yu J. (1990). The inhibin/activin family of hormones and growth factors. In: Peptide growth factors and their receptors n. Spom MB, Roberts AB, editors. Berlin: Springer-Verlag, pp. 211-248.
• Vassalli A., Matzuk MM, Gardner HA, Lee KF, Jaenisch R. (1994). Activin/inhibin βB subunit gene disruption leads to defects in eyelid development and female reproduction. Genes Dev 8:414-427.[Abstract/Free Full Text]
• Verschueren K., Dewulf N., Goumans M-J., Lonnoy O., Feijen A., Grimsby S., et al. (1995). Expression of type I and type IB receptors for activin in midgestation mouse embryos suggests distinct functions in organogenesis. Mech Dev 52:109-123.[CrossRef][Medline] [Order article via Infotrieve]

Journal of Dental Research, Vol. 76, No. 10, 1625-1636 (1997)
DOI: 10.1177/00220345970760100301


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				H. Lesot, S. Lisi, R. Peterkova, M. Peterka, V. Mitolo, and J.V. Ruch Epigenetic Signals during Odontoblast Differentiation Advances in Dental Research, August 1, 2001; 15(1): 8 - 13. [Abstract] [PDF]

				A.J. Smith and H. Lesot Induction and Regulation of Crown Dentinogenesis: Embryonic Events as a Template for Dental Tissue Repair? Critical Reviews in Oral Biology & Medicine, January 1, 2001; 12(5): 425 - 437. [Abstract] [Full Text] [PDF]

				C. A. Ferguson, A. S. Tucker, L. Christensen, A. L. Lau, M. M. Matzuk, and P. T. Sharpe Activin is an essential early mesenchymal signal in tooth development that is required for patterning of the murine dentition Genes & Dev., August 15, 1998; 12(16): 2636 - 2649. [Abstract] [Full Text]

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Heikinheimo, K. Articles by Ruch, J.V. Search for Related Content PubMed PubMed Citation Articles by Heikinheimo, K. Articles by Ruch, J.V. Social Bookmarking                         What's this?