Skip to main content
Log in

The Distribution of BrdU- and TUNEL-positive Cells During Odontogenesis in Mouse Lower First Molars

  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Abstract

This study investigated the minute distribution of both proliferating and non-proliferating cells, and cell death in the developing mouse lower first molars using 5-bromo-2′-deoxyuridine (BrdU) incorporation and the terminal deoxynucleotidyl transferase-mediated deoxyuridine-5′-triphosphate (dUTP)-biotin nick end labeling (TUNEL) double-staining technique. The distribution pattern of the TUNEL-positive cells was more notable than that of the BrdU-positive cells. TUNEL-positive cells were localized in the following six sites: (1) in the most superficial layer of the dental epithelium during the initiation stage, (2) in the dental lamina throughout the period during which tooth germs grow after bud formation, (3) in the dental epithelium in the most anterior part of the antero-posterior axis of the tooth germ after bud formation, (4) in the primary enamel knot from the late bud stage to the late cap stage, (5) in the secondary enamel knots from the late cap stage to the late bell stage, and (6) in the stellate reticulum around the tips of the prospective cusps after the early bell stage. These peculiar distributions of TUNEL-positive cells seemed to have some effect on either the determination of the exact position of the tooth germ in the mandible or on the complicated morphogenesis of the cusps. The distribution of BrdU-negative cells was closely associated with TUNEL-positive cells, which thus suggested cell arrest and the cell death to be essential for the tooth morphogenesis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References cited

  • Aberg T, Wozney J, Thesleff I (1997) Expression patterns of bone morphogenetic proteins (Bmps) in the developing mouse tooth suggest roles in morphogenesis and cell differentiation. Dev Dyn 210: 383-396.

    Google Scholar 

  • Alison MR (1992) Repair and regenerative responses. In McGee JO'D, Issacson PG, Wright NA, ed. Oxford Textbook of Pathology. Oxford: Oxford University Press, pp. 365-379.

    Google Scholar 

  • Ayer-Le Lievre C, Stahlbom PA, Sara VR (1991) Expression of IGF-I and-II mRNA in the brain and craniofacial region of the rat fetus. Development 111: 105-115.

    Google Scholar 

  • Cam Y, Neumann MR, Oliver L, Raulais D, Janet T, Ruch JV (1992) Immunolocalization of acidic and basic fibroblast growth factors during mouse odontogenesis. Int J Dev Biol 36: 381-389.

    Google Scholar 

  • Casasco A, Calligaro A, Casasco M (1992) Proliferative and functional stages of rat ameloblast differentiation as revealed by combined immunocytochemistry against enamel matrix proteins and bromodeoxyuridine. Cell Tissue Res 270: 415-423.

    Google Scholar 

  • Casasco A, Casasco M, Cornaglia AI, Mazzini G, De Renzis R, Tateo S (1995) Detection of bromo-deoxyuridine-and proliferating cell nuclear antigen-immunoreactivities in tooth germ. Connect Tissue Res 32: 63-70.

    Google Scholar 

  • Cohn MJ, Izpisua-Belmonte JC, Abud H, Heath JK, Tickle C (1995) Fibroblast growth factors induce additional limb development from the flank of chick embryos. Cell 80: 739-746.

    Google Scholar 

  • Gavrieli Y, Sherman Y, Ben-Sasson SA (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119: 493-501.

    Google Scholar 

  • Graham A, Francis-West P, Brickell P, Lumsden A (1994) The signalling molecule BMP4 mediates apoptosis in the rhombencephalic neural crest. Nature 372: 684-686.

    Google Scholar 

  • Graham A, Koentges G, Lumsden A (1996) Neural crest apoptosis and the establishment of craniofacial pattern: an honorable death. Mol Cell Neurosci 8: 76-83.

    Google Scholar 

  • Hermeking H, Eick D (1994) Mediation of c-Myc-induced apoptosis by p53. Science 265: 2091-2093.

    Google Scholar 

  • Jernvall J, Kettunen P, Karavanova I, Martin LB, Thesleff I (1994) Evidence for the role of the enamel knot as a control center in mammalian tooth cusp formation: non-dividing cells express growth stimulating Fgf-4 gene. Int J Dev Biol 38: 463-469.

    Google Scholar 

  • Jernvall J (1995) Mammalian molar cusp patterns: developmental mechanisms of diversity. Acta Zool Fennica 198: 1-61.

    Google Scholar 

  • Jernvall J, Aberg T, Kettunen P, Keranen S, Thesleff I (1998) The life history of an embryonic signaling center: BMP-4 induces p21 and is associated with apoptosis in the mouse tooth enamel knot. Development 125: 161-169.

    Google Scholar 

  • Jowett AK, Vainio S, Ferguson MW, Sharpe PT, Thesleff I (1993) Epithelial-mesenchymal interactions are required for msx 1 and msx 2 gene expression in the developing murine molar tooth. Development 117: 461-470.

    Google Scholar 

  • Kaartinen V, Cui XM, Heisterkamp N, Groffen J, Shuler CF (1997) Transforming growth factor-beta3 regulates transdifferentiation of medial edge epithelium during palatal fusion and associated degradation of the basement membrane. Dev Dyn 209: 255-260.

    Google Scholar 

  • Krajewski S, Hugger A, Krajewska M, John C. Reed JC, Mai JK (1998) Developmental expression patterns of Bcl-2, Bcl-x, Bax, and Bak in teeth. Cell Death Differ 5: 408-415.

    Google Scholar 

  • Lesot H, Vonesch JL, Peterka M, Tureckova J, Peterkova R, Ruch JV (1996) Mouse molar morphogenesis revisited by three-dimensional reconstruction. II. Spatial distribution of mitoses and apoptosis in cap to bell staged first and second upper molar teeth. Int J Dev Biol 40: 1017-1031.

    Google Scholar 

  • Maas R, Bei M (1997) The genetic control of early tooth development. Crit Rev Oral Biol Med 8: 4-39.

    Google Scholar 

  • Macias D, Ganan Y, Sampath TK, Piedra ME, Ros MA, Hurle JM (1997) Role of BMP-2 and OP-1 (BMP-7) in programmed cell death and skeletogenesis during chick limb development. Development 124: 1109-1117.

    Google Scholar 

  • Mackenzie A, Ferguson MW, Sharpe PT (1992) Expression patterns of the homeobox gene, Hox-8, in the mouse embryo suggest a role in specifying tooth initiation and shape. Development 115: 403-420.

    Google Scholar 

  • Marazzi G, Wang Y, Sassoon D (1997) Msx2 is a transcriptional regulator in the BMP4-mediated programmed cell death pathway. Dev Biol 186: 127-138.

    Google Scholar 

  • Matsumoto K, Nakamura T (1997) Hepatocyte growth factor (HGF) as a tissue organizer for organogenesis and regeneration. Biochem Biophys Res Commun 239: 639-644.

    Google Scholar 

  • Mitsiadis TA, Salmivirta M, Muramatsu T, Muramatsu H, Rauvala H, Lehtonen E, Jalkanen M, Thesleff I (1995a) Expression of the heparinbinding cytokines, midkine (MK) and HB-GAM (pleiotrophin) is associated with epithelial-mesenchymal interactions during fetal development and organogenesis. Development 121: 37-51.

    Google Scholar 

  • Mitsiadis TA, Muramatsu T, Muramatsu H, Thesleff I (1995b) Midkine (MK), a heparin-binding growth/differentiation factor, is regulated by retinoic acid and epithelial-mesenchymal interactions in the developing mouse tooth, and affects cell proliferation and morphogenesis. J Cell Biol 129: 267-281.

    Google Scholar 

  • Niswander L, Martin GR (1993) FGF-4 and BMP-2 have opposite effects on limb growth. Nature 361: 68-71.

    Google Scholar 

  • Niswander L, Tickle C, Vogel A, Booth I, Martin GR (1993) FGF-4 replaces the apical ectodermal ridge and directs outgrowth and patterning of the limb. Cell 75: 579-587.

    Google Scholar 

  • Parker SB, Eichele G, Zhang P, Rawls A, Sands AT Bradley A, Olson EN, Harper JW, Elledge SJ (1995) p53-independent expression of p21Cip1 in muscle and other terminally differentiating cells. Science 267: 1024-1027.

    Google Scholar 

  • Pelton RW, Dickinson ME, Moses HL, Hogan BL (1990) In situ hybridization analysis of TGF beta 3 RNA expression during mouse development: comparative studies with TGF beta 1 and beta 2. Development 110: 609-620.

    Google Scholar 

  • Ros MA, Piedra ME, Fallon JF, Hurle JM (1997) Morphogenetic potential of the chick leg interdigital mesoderm when diverted from the cell death program. Dev Dyn 208: 406-419.

    Google Scholar 

  • Ruch JV (1995) Tooth crown morphogenesis and cytodifferentiations: candid questions and critical comments. Connect Tissue Res 32: 1-8.

    Google Scholar 

  • Slootweg PJ, De Weger RA (1994) Immunohistochemical demonstration of bcl-2 protein in human tooth germs. Arch Oral Biol 39: 545-550.

    Google Scholar 

  • Snead ML, Luo W, Oliver P, Nakamura M, Don-Wheeler G, Bessem C, Bell GI, Rall LB, Slavkin HC (1989) Localization of epidermal growth factor precursor in tooth and lung during embryonic mouse development. Dev Biol 134: 420-429.

    Google Scholar 

  • Sonnenberg E, Weidner KM, Birchmeier C (1993) Expression of the met-receptor and its ligand, HGF-SF during mouse embryogenesis. EXS 65: 381-394.

    Google Scholar 

  • Tabata MJ, Kim K, Liu JG, Yamashita K, Matsumura T, Kato J, Iwamoto M, Wakisaka S, Matsumoto K, Nakamura T, Kumegawa M, Kurisu K (1996) Hepatocyte growth factor is involved in the morphogenesis of tooth germ in murine molars. Development 122: 1243-1251.

    Google Scholar 

  • Ten Cate AR (1994) Development of the tooth and its supporting tissue. In Oral Histology. Development, Structure and Function. St Louis: Mosby, pp. 58-80.

    Google Scholar 

  • Thesleff I, Nieminen P (1996) Tooth morphogenesis and cell differentiation. Curr Opin Cell Biol 8: 844-850.

    Google Scholar 

  • Vaahtokari A, Vainio S, Thesleff I (1991) Associations between transforming growth factor beta 1 RNA expression and epithelial-mesenchymal interactions during tooth morphogenesis. Development 113: 985-994.

    Google Scholar 

  • Vaahtokari A, Aberg T, Jernvall J, Keranen S, Thesleff I (1996a) The enamel knot as a signaling center in the developing mouse tooth. Mech Dev 54: 39-43.

    Google Scholar 

  • Vaahtokari A, Aberg T, Thesleff I (1996b) Apoptosis in the developing tooth: association with an embryonic signaling center and suppression by EGF and FGF-4. Development 122: 121-129.

    Google Scholar 

  • Vainio S, Thesleff I (1992) Sequential induction of syndecan, tenascin and cell proliferation associated with mesenchymal cell condensation during early tooth development. Differentiation 50: 97-105.

    Google Scholar 

  • 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 early tooth development. Cell 75: 45-58.

    Google Scholar 

  • Vaux DL, Cory S, Adams JM (1988) Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cells. Nature 335: 440-442.

    Google Scholar 

  • Viriot L, Peterkova R, Vonesch JL, Peterka M, Ruch JV, Lesot H (1997) Mouse molar morphogenesis revisited by three-dimensional reconstruction. III. Spatial distribution of mitoses and apoptoses up to bellstaged first lower molar teeth. Int J Dev Biol 41: 679-690.

    Google Scholar 

  • Wagner AJ, Kokontis JM, Hay N (1994) Myc-mediated apoptosis requires wild-type p53 in a manner independent of cell cycle arrest and the ability of p53 to induce p21waf1/cip1. Genes Dev 8: 2817-2830.

    Google Scholar 

  • Werner H, Le Roith D (1997) The insulin-like growth factor-I receptor signaling pathways are important for tumorigenesis and inhibition of apoptosis. Crit Rev Oncog 8: 71-92.

    Google Scholar 

  • Xu RH, Dong Z, Maeno M, Kim J, Suzuki A, Ueno N, Sredni D, Colburn NH, Kung HF (1996) Involvement of Ras/Raf/AP-1 in BMP-4 signaling during Xenopus embryonic development. Proc Natl Acad Sci USA 93: 834-838.

    Google Scholar 

  • Young WG, Ruch JV, Stevens MR, Begue-Kirn C, Zhang CZ, Lesot H, Waters MJ (1995) Comparison of the effects of growth hormone, insulin-like growth factor-I and fetal calf serum on mouse molar odontogenesis in vitro. Arch Oral Biol 40: 789-799.

    Google Scholar 

  • Zou H, Niswander L (1996) Requirement for BMP signaling in interdigital apoptosis and scale formation. Science 272: 738-741.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shigemura, N., Kiyoshima, T., Kobayashi, I. et al. The Distribution of BrdU- and TUNEL-positive Cells During Odontogenesis in Mouse Lower First Molars. Histochem J 31, 367–377 (1999). https://doi.org/10.1023/A:1003796023992

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1003796023992

Keywords

Navigation