Skip to main content
SpringerLink
Log in

Gap junction proteins exhibit early and specific expression during intramembranous bone formation in the developing chick mandible

  • Original Articles
  • Published:
Anatomy and Embryology Aims and scope Submit manuscript

Abstract

The spatial and temporal expression of three closely related members of the connexin family of gap junction proteins (connexin42, Cx42; connexin43, Cx43; and connexin45, Cx45) was evaluated during bone formation in the mandibular process of the chick embryo. Mandibles of chick embryos from Hamburger and Hamilton stage 25 (approximately 5 days) through 19 days of development were dissected, serially sectioned and processed for immunocytochemical localization, employing site-specific anti-connexin antibodies. Our data revealed that (1) Cx43 was present throughout mandibular bone formation; (2) although it appeared to be associated with all bone cell types, Cx43 was concentrated in mesenchymal cells during the earliest stages in the osteogenic lineage; (3) most importantly, the localization of Cx43 at sites of bone formation appeared to precede the overt expression of the osteogenic phenotype; (4) by contrast, Cx45 was more restricted, spatially and temporally, in its distribution; (5) Cx42 expression was not detected in osteogenic tissue during mandibular bone formation. From all of the data obtained, Cx45 appeared to be associated with stages of bone formation characterized by the elaboration of matrix and the progressive expression of the differentiated osteogenic phenotype. Cx43 appeared to be associated with condensation of mesenchyme and the earliest stages of osteogenesis. Because of these associations, we propose that connexin expression may be necessary for the initiation of bone formation and the full expression of the osteogenic phenotype.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Beyer EC (1990) Molecular cloning and developmental expression of two chick embryo gap junction proteins. J Biol Chem 265:14439–14443

    CAS  PubMed  Google Scholar 

  • Bhargava U, Bar-lev M, Bellows CG, Aubin JE (1988) Ultrastructural analysis of bone nodules formed in vitro by isolated fetal rat calvaria cells. Bone 9:155–163

    Article  CAS  PubMed  Google Scholar 

  • Bruder SP, Caplan AI (1989) First bone formation and the dissection of an osteogenic lineage in the embryonic chick tibia is revealed by monoclonal antibodies against osteoblasts. Bone 10:359–375

    Google Scholar 

  • Bruder SP, Caplan AI (1990a) A monoclonal antibody against the surface of osteoblasts recognizes alkaline phosphatase isoenzymes in bone, liver, kidney, and intestine. Bone 11:133–139

    Google Scholar 

  • Bruder SP, Caplan AI (1990b) Terminal differentiation of osteogenic cells in the embryonic chick tibia is revealed by a monoclonal antibody against osteocytes. Bone 11:189–198

    Google Scholar 

  • Civitelli R, Beyer EC, Warlow PM, Robertson AJ, Geist ST, Steinberg TH (1993) Connexin43 mediates direct intercellular communication in human osteoblastic cell networks. J Clin Invest 91:1888–1896

    Google Scholar 

  • Coelho CND, Kosher RA (1991) Gap junctional communication during limb cartilage differentiation. Dev Biol 144:47–53

    Google Scholar 

  • Doty SB (1981) Morphological evidence of gap junctions between bone cells. Calcif Tissue Int 33:509–512

    Google Scholar 

  • Doty SB (1988) Cell-to-cell communication in bone tissue. In: Davidovitch Z (ed) The biological mechanisms of tooth eruption and root resorption. EBSCO Media, Birmingham, Ala, USA, pp 61–69

    Google Scholar 

  • Gerstenfeld LC, Gotoh Y, McKee MD, Nanci A, Landis WJ, Glimher MJ (1990) Expression and ultrastructural immunolocalization of a major 66-kDa phosphoprotein synthesized by chicken osteoblast during mineralization in vitro. Anat Rec 228:93–103

    Google Scholar 

  • Gotoh Y, Gerstenfeld LC, Glimcher MJ (1990) Identification and characterization of the major chicken bone phosphoprotein: analysis of its synthesis by cultured embryonic chick osteoblasts. Eur J Biochem 187:49–58

    Google Scholar 

  • Hall BK (1978) Initiation of osteogenesis by mandibular mesenchyme of the embryonic chick in response to mandibular and non-mandibular epithelia. Arch Oral Biol 23:1157–1161

    Google Scholar 

  • Hall BK (1980) Viability and proliferation of epithelia and the initiation of osteogenesis within mandibular ectomesenchyme in the embryonic chick. J Embryol Exp Morphol 56:71–89

    Google Scholar 

  • Hall BK (1983) Epithelial-mesenchymal interactions in cartilage and bone development. In: Sawyer RH, Follon JF (eds) Epithelial-mesenchymal interactions in development. Praeger, New York, pp 189–214

    Google Scholar 

  • Hall BK, Miyake T (1992) The membranous skeleton: the role of cell condensations in vertebrate skeletogenesis. Anat Embryol 186:107–124

    Google Scholar 

  • Hall BK, Van Exan RJ (1982) Induction of bone by epithelial cell products. J Embryol Exp Morphol 69:37–46

    Google Scholar 

  • Hamburger V, Hamilton H (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–92

    Google Scholar 

  • Henneman H, Schwarz HJ, Willecke K (1992) Characterization of gap junction genes expressed in F9 embryonic carcinoma cells: molecular cloning of mouse connexin31 and -45 cDNAs. Eur J Cell Biol 57:51–58

    Google Scholar 

  • Jones SJ, Gray C, Sakamaki H, Arora M, Boyde A, Gourdie R, Green C (1993) The incidence and size of gap junctions between the bone cells in rat calvaria. Anat Embryol 187:343–352

    Google Scholar 

  • Jongen WMF, Fitzgerald DJ, Asamoto M, Piccoli C, Slaga TJ, Gros D, Takeichi M, Yamasaki H (1991) Regulation of connex-in43-mediated gap junctional intercellular communication by Ca2+ in mouse epidermal cells is controlled by Ecadherin. J Cell Biol 114:545–555

    Google Scholar 

  • Kanter HL, Saffitz JE, Beyer EC (1992) Cardiac myocytes express multiple gap junction proteins. Circ Res 70:438–444

    Google Scholar 

  • Kanter HL, Laing JG, Beyer EC, Green KG, Saffitz JE (1993) Multiple connexins colocalize in canine ventricular myocyte gap junctions. Circ Res 73:344–350

    Google Scholar 

  • Keane RW, Mehta PP, Rose B, Honig LS, Loewenstein WR, Rutishauser U (1988) Neural differentiation, NCAM-mediated adhesion, and gap junctional communication in neuroectoderm. A study in vitro. J Cell Biol 106:1307–1319

    Google Scholar 

  • Le Lièvre C (1978) Participation of neural crest-derived cells in the genesis of the skull in birds. J Embryol Exp Morphol 47:17–37

    Google Scholar 

  • Matsuzaki F, Mège R-M, Jaffe SH, Friedlander DR, Gallin WJ, Goldberg JI, Cunningham BA, Edelman GM (1990) cDNAs of cell adhesion molecules of different specificity induce changes in cell shape and border formation in cultured S180 cells. J Cell Biol 110:1239–1252

    Google Scholar 

  • McKee MD, Nanci A, Landis WJ, Gotoh Y Gerstenfeld LC, Glimcher MJ (1990) Developmental appearance and ultrastructural immunolocalization of a major 66 kDa phosphoprotein in embryonic and post-natal chicken bone. Anat Rec 228:77–92

    Google Scholar 

  • Mège R-M, Matsuzaki F, Gallin WJ, Goldberg JI, Cunningham BA, Edelman GM (1988) Construction of epithelioid sheets by transfection of mouse sarcoma cells with cDNAs for chicken cell adhesion molecules. Proc Natl Acad Sci USA 85:7274–7278

    Google Scholar 

  • Meyer RA, Laird DW, Revel J-P, Johnson RG (1992) Inhibition of gap junction and adherens junction assembly by connexin and A-CAM antibodies. J Cell Biol 119:179–189

    Google Scholar 

  • Miller SC, Saint-Georges L de, Bowman BM, Jee WSS (1989) Bone lining cells: structure and function. Scanning Microsc 3:953–961

    Google Scholar 

  • Mina M, Kollar EJ, Upholt WB (1991) Temporal and spatial expression of genes for cartilage extracellular matrix proteins during avian mandibular arch development. Differentiation 48:17–24

    Google Scholar 

  • Minkoff R, Parker SB, Hertzberg EL (1991) Analysis of distribution patterns of gap junctions during development of embryonic chick facial primordia and brain. Development 111:509–522

    Google Scholar 

  • Minkoff R, Parker SB, Neuman PN, Hertzberg EL (1992) Spatiotemporal expression of gap junctions during primary palate closure (abstract). J Dent Res 71:683

    Google Scholar 

  • Minkoff R, Rundus VR, Parker SB, Beyer EC, Hertzberg EL (1993) Connexin expression in the developing avian cardiovascular system. Circ Res 73:71–78

    Google Scholar 

  • Murray PDF (1963) Adventitious (secondary) cartilage in the chick embryo and the development of certain bones and articulations in the chick skull. Aust J Zool 11:368–430

    Google Scholar 

  • Musil LS, Beyer EC, Goodenough DA (1990a) Expression of the gap junction protein connexin43 in embryonic chick lens: molecular cloning, ultrastructural localization, and post-translational phosphorylation. J Membr Biol 116:163–175

    CAS  PubMed  Google Scholar 

  • Musil LS, Cunningham BA, Edelman GM, Goodenough DA (1990b) Differential phosphorylation of the gap junction protein connexin43 in junctional communication-competent and -deficient cell lines. J Cell Biol 111:2077–2088

    Google Scholar 

  • Nishi M, Kumar NM, Gilula NB (1991) Developmental regulation of gap junction gene expression during mouse embryonic development. Dev Biol 146:117–130

    Google Scholar 

  • Noden DM (1978) The control of avian cephalic neural crest cytodifferentiation. I. Skeletal and connective tissues. Dev Biol 67:296–312

    Google Scholar 

  • Noden DM (1980) The migration and cytodifferentiation of cranial neural crest cells. In: Pratt RM, Christiansen RL (eds) Current research trends in prenatal craniofacial development. Elsevier/North Holland, New York, pp 3–25

    Google Scholar 

  • Noden DM (1991) Cell movements and control of patterned tissue assembly during craniofacial development. J Craniofac Genet Dev Biol 11:192–213

    Google Scholar 

  • Parker SB, Butler WT, Hertzberg EL, Minkoff R (1992) Expression of gap junctions in osteogenic tissue during mandibular development (abstract). J Dent Res 71:683

    Google Scholar 

  • Risek BK, Klier FG, Gilula NB (1992) Multiple gap junction genes are utilized during rat skin and hair development. Development 116:639–651

    Google Scholar 

  • Ruangvoravat CP, Lo CW (1992) Connexin 43 expression in the mouse embryo: localization of transcripts within developmentally significant domains. Dev Dyn 194:261–281

    CAS  PubMed  Google Scholar 

  • Sainio K, Gilbert SF, Lehtonen E, Nishi M, Kumar NM, Gilula NB, Saxen L (1992) Differential expression of gap junction mRNAs and proteins in the developing murine kidney and in experimentally induced nephric mesenchymes. Development 115:827–837

    Google Scholar 

  • Schiller PC, Mehta PP, Roos BA, Howard GA (1992) Hormonal regulation of intercellular communication: parathyroid hormone increases connexin 43 gene expression and gap-junctional communication in osteoblastic cells. Mol Endocrinol 6:1433–1440

    Google Scholar 

  • Schirrmacher K, Schmitz I, Winterhager E, Traub O, Brümmer F, Jones D, Bingmann D (1992) Characterization of gap junctions between osteoblast-like cells in culture. Calcif Tissue Int 51:285–290

    Google Scholar 

  • Shapiro F (1988) Cortical bone repair. J Bone Joint Surg 70:1067–1081

    Google Scholar 

  • Shen V, Rifas E, Kohler G, Peck WA (1986) Prostaglandins change cell shape and increase intercellular gap junctions in osteoblasts cultured from rat fetal calvaria. J Bone Min Res 1:243–249

    Google Scholar 

  • Takahashi Y, Bontoux M, Le Douarin NM (1991) Epithelio-mesenchymal interactions are critical for Quox 7 expression and membrane bone differentiation in the neural crest-derived mandibular mesenchyme. Embo J 10:2387–2393

    Google Scholar 

  • Tyler MS, Hall BK (1977) Epithelial influences on skeletenogenesis in the mandible of the embryonic chick. Anat Rec 188:229–240

    Google Scholar 

  • Warner A (1992) Gap junctions in development — a perspective. Semin Cell Biol 3:81–91

    Google Scholar 

  • Yamamoto T, Ochalski A, Hertzberg EL, Nagy JI (1990) LM and EM immunolocalization of the gap junctional connexin 43 in rat brain. Brain Res 508:313–319

    Google Scholar 

  • Yamamoto T, Hossain MZ, Hertzberg EE, Uemura H, Murphy LJ, Nagy JI (1993) Connexin43 in rat pituitary: localization at pituicyte and stellate cell gap junctions and within gonadotrophs. Histochemistry 100:53–64

    Google Scholar 

  • Yancey SB, Biswal S, Revel JP (1992) Spatial and temporal patterns of distribution of the gap junction protein connexin43 during mouse gastrulation and organogenesis. Development 114:203–212

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Craniofacial Growth and Development, Dental Branch, University of Texas, Health Science Center at Houston, 77225, Houston, TX, USA

    R. Minkoff, V. R. Rundus & S. B. Parker

  2. Department of Neuroscience and Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, 10461, Bronx, NY, USA

    E. L. Hertzberg

  3. Department of Pediatrics, Medicine, and Cell Biology and Physiology, Washington University School of Medicine, 63110, St. Louis, MO, USA

    J. G. Laing & E. C. Beyer

Authors
  1. R. Minkoff
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. V. R. Rundus
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. B. Parker
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. E. L. Hertzberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J. G. Laing
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. E. C. Beyer
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Minkoff, R., Rundus, V.R., Parker, S.B. et al. Gap junction proteins exhibit early and specific expression during intramembranous bone formation in the developing chick mandible. Anat Embryol 190, 231–241 (1994). https://doi.org/10.1007/BF00234301

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00234301

Key words

Search

Navigation