Skip to main content

Advertisement

SpringerLink
Log in

Adhesion structures and their cytoskeleton-membrane interactions at podosomes of osteoclasts in culture

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

The organization of the cytoskeleton in the podosomes of osteoclasts was studied by use of cell shearing, rotary replication, and fluorescence cytochemical techniques. After shearing, clathrin plaques and particles associated with the cytoskeleton were left behind on the exposed cytoplasmic side of the membrane. The cytoskeleton of the podosomes was characterized by two types of actin filaments: relatively long filaments in the portion surrounding the podosome core, and highly branched short filaments in the core. Individual actin filaments radiating from the podosomes interacted with several membrane particles along the length of the filaments. Many lateral contacts with the membrane surface by the particles were made along the length of individual actin filaments. The polarity of actin filaments in podosomes became oriented such that their barbed ends were directed toward the core of podosomes. The actin cytoskeletons terminated or branched at the podosomes, where the membrane tightly adhered to the substratum. Microtubules were not usually present in the podosome structures; however, certain microtubules appeared to be morphologically in direct contact with the podosome core. Most of the larger clathrin plaques consisted of flat sheets of clathrin lattices that interconnected neighboring clathrin lattices to form an extensive clathrin area. However, the small deeply invaginated clathrin plaques and the podosomal cytoskeleton were located close together. Thus, the clathrin plaques on the ventral membrane of osteoclasts might be involved in both cell adhesion and the formation of receptor-ligand complexes, i.e., endocytosis.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12
Fig. 13

Similar content being viewed by others

References

  • Aggeler J, Takemura R, Werb Z (1983) High-resolution three-dimensional views of membrane-associated clathrin and cytoskeleton in critical-point-dried macrophages. J Cell Biol 97:1452–1458

    Article  PubMed  CAS  Google Scholar 

  • Akisaka T, Miyaji T, Yoshida H, Inoue M (1997) Ultrastructure of quick-frozen and freeze-substituted chick osteoclasts. J Anat 190:433–445

    Article  PubMed  Google Scholar 

  • Akisaka T, Yoshida H, Inoue S, Shimizu K (2001) Organization of cytoskeletal F-actin, G-actin, and gelsolin in the adhesion structures in cultured osteoclast. J Bone Miner Res 16:1248–1255

    Article  PubMed  CAS  Google Scholar 

  • Akisaka T, Yoshida H, Suzuki R, Shimizu K, Takama K (2003) Clathrin sheets on the protoplasmic surface of ventral membranes of osteoclasts in culture. J Electron Microsc 52:535–543

    Article  CAS  Google Scholar 

  • Akisaka T, Yoshida H, Suzuki R (2006) The ruffled border and attachment regions of the apposing membrane of resorbing osteoclasts as visualized from the cytoplasmic face of the membrane. J Electron Microsc 55:53–61

    Article  CAS  Google Scholar 

  • Aubin JE (1992) Osteoclast adhesion and resorption: the role of podosomes. J Bone Miner Res 7:365–368

    PubMed  CAS  Google Scholar 

  • Avnur Z, Geiger B (1981) Substrate-attached membranes of cultured cells: isolation and characterization of ventral cell membranes and the associated cytoskeleton. J Mol Biol 153:361–379

    Article  PubMed  CAS  Google Scholar 

  • Babb SG, Matsudaira P, Sato M, Correia I, Lim SS (1997) Fimbrin in podosomes of monocyte-derived osteoclasts. Cell Motil Cytoskelet 37:308–325

    Article  CAS  Google Scholar 

  • Buccione R, Orth JD, McNiven MA (2004) Foot and mouth: podosomes, invadopodia and circular dorsal ruffles. Nat Rev Mol Cell Biol 5:647–657

    Article  PubMed  CAS  Google Scholar 

  • Burns S, Thrasher AJ, Blundell TM, Machesky L, Jones GE (2001) Configuration of human dendritic cell cytoskeleton by Rho GTPases, the WAS protein, and differentiation. Blood 98:1142–1149

    Article  PubMed  CAS  Google Scholar 

  • Cooper JA (1991) The role of actin polymerization in cell motility. Annu Rev Physiol 53:585–605

    Article  PubMed  CAS  Google Scholar 

  • Correia I, Chu D, Chou Y-H, Goldman RD, Matsudaira P (1999) Integrating the actin and vimentin cytoskeletons: adhesion-dependent formation of fimbrin-vimentin complexes in macrophages. J Cell Biol 146:831–842

    Article  PubMed  CAS  Google Scholar 

  • David-Pfeuty T, Singer SJ (1980)Altered distributions of the cytoskeletal proteins vinculin and alpha-actinin in cultured fibroblasts transformed by Rous sarcoma virus. Proc Natl Acad Sci USA 77:6687–6691

    Article  PubMed  CAS  Google Scholar 

  • Destaing O, Saltel F, Geminard J-C, Jurdic P, Bard F (2003) Podosomes display actin turnover and dynamic self-organization in osteoclasts expressing actin-green fluorescent protein. Mol Biol Cell 14:407–416

    Article  PubMed  CAS  Google Scholar 

  • Destaing O, Saltel F, Gilquin B, Chabadel A, Khochbin S, Ory S, Jurdic P (2005) A novel Rho-mDia2-HDAC6 pathway controls podosome patterning through microtubule acetylation in osteoclasts. J Cell Sci 118:2901–2911

    Article  PubMed  CAS  Google Scholar 

  • Evans JG, Matsudaira P (2006) Structure and dynamics of macrophage podosomes. Eur J Cell Biol 85:145–149

    Article  PubMed  CAS  Google Scholar 

  • Evans JG, Correia I, Krasavina O, Watson N, Matsudaira P (2003) Macrophage podosomes assemble at the leading lamella by growth and fragmentation. J Cell Biol 161:697–705

    Article  PubMed  CAS  Google Scholar 

  • Felice GR, Eason P, Nermut MV, Kellie S (1990) pp60v-src association with the cytoskeleton induces actin reorganization without affecting polymerization status. Eur J Cell Biol 52:47–59

    PubMed  CAS  Google Scholar 

  • Gavazzi I, Nermut MV, Marchisio PC (1989) Ultrastructure and gold-immunolabelling of cell-substratum adhesions (podosomes) in RSV-transformed BHK cells. J Cell Sci 94:85–99

    PubMed  Google Scholar 

  • Goode BL, Drubin DG, Barness G (2000) Functional cooperation between the microtubule and actin cytoskeletons. Curr Opin Cell Biol 12:63–71

    Article  PubMed  CAS  Google Scholar 

  • Heuser JE, Anderson RGW (1989) Hypertonic media inhibit receptor-mediated endocytosis by blocking clathrin-coated pit formation. J Cell Biol 108:389–400

    Article  PubMed  CAS  Google Scholar 

  • Heuser JE, Cooke R (1983) Actin-myosin interactions visualized by the quick-freeze, deep-etch replica technique. J Mol Biol 169:97–122

    Article  PubMed  CAS  Google Scholar 

  • Jurdic P, Saltel F, Chabadel A, Destaing O (2006) Podosome and sealing zone: specificity of the osteoclast model. Eur J Cell Biol 85:195–202

    Article  PubMed  CAS  Google Scholar 

  • Kaverina I, Krylyshkina O, Small JV (1999) Microtubule targeting of substrate contacts promotes their relaxation and dissociation. J Cell Biol 146:1033–1044

    Article  PubMed  CAS  Google Scholar 

  • Kaverina I, Krylyshkina O, Small JV (2002) Regulation of substrate adhesion dynamics during cell motility. Int J Biochem Cell Biol 34:746–761

    Article  PubMed  CAS  Google Scholar 

  • Kaverina I, Stradal TEB, Gimona M (2003) Podosome formation in cultures A7r5 vascular smooth muscle cells requires Arp2/3-dependent de-novo actin polymerization at discrete microdomains. J Cell Biol 116:4915–4924

    CAS  Google Scholar 

  • Keating TJ, Peloquin JG, Rodionov VI, Momcilovic D, Borisy GG (1997) Microtubule release from the centrosome. Proc Natl Acad Sci USA 94:5078–5083

    Article  PubMed  CAS  Google Scholar 

  • Lakkakorpi P, Tuukkanen J, Hentunen T, Jarvelin K, Vaananen HK (1989) Organization of osteoclast microfilaments during the attachment to bone surface in vitro. J Bone Miner Res 4:817–825

    Article  PubMed  CAS  Google Scholar 

  • Lakkakorpi PT, Helfrich MH, Horton MA, Vaananen HK (1993) Spatial organization of microfilaments and vitronectin receptor, alpha v beta 3, in osteoclasts. J Cell Sci 104:663–670

    PubMed  CAS  Google Scholar 

  • Larkin JM, Donzell WC, Anderson RGW (1986) Potassium-dependent assembly of coated pits: new coated pits form as planar clathrin lattices. J Cell Biol 103:2619–2627

    Article  PubMed  CAS  Google Scholar 

  • Lewis AK, Bridgman PC (1992) Nerve growth cone lamellipodia contain two populations of actin filaments that differ in organization and porality. J Cell Biol 119:1219–1243

    Article  PubMed  CAS  Google Scholar 

  • Linder S, Aepfelbacher M (2003) Podosomes: adhesion hot-spots of invasive cells. Trends Cell Biol 13:376–385

    Article  PubMed  CAS  Google Scholar 

  • Linder S, Kopp P (2005) Podosomes at a glance. J Cell Sci 118:2079–2082

    Article  PubMed  CAS  Google Scholar 

  • Linder S, Higgs H, Hufner K, Schwarz K, Pannicke U, Aepfelbacher M (2000a) The polarization defect of Wiskott-Aldrich syndrome macrophages is linked to dislocalization of the Arp2/3 complex. J Immunol 165:221–225

    PubMed  CAS  Google Scholar 

  • Linder S, Hufner K, Wintergerst U, Apefelbacher M (2000b) Microtubule-dependent formation of podosomal adhesion structures in primary human macrophages. J Cell Sci 113:4165–4176

    PubMed  CAS  Google Scholar 

  • Luther PW, Samuelsson SJ, Bloch RJ, Pumplin DW (1996) Ctoskeleton-membrane interactions at the postsynaptic density of Xenopus neuromuscular junctions. J Neurocytol 25:417–427

    PubMed  CAS  Google Scholar 

  • Luxenburg C, Addadi L, Geiger B (2006) The molecular dynamics of osteoclast adhesions. Eur J Cell Biol 85:203–211

    Article  PubMed  CAS  Google Scholar 

  • Luxenburg C, Geblinger D, Klein E, Anderson K, Hanein D, Geiger B, Addadi L (2007) The architecture of the adhesive apparatus of cultured osteoclasts: from podosome formation to sealing zone assembly. PLos One 2:e179

    Article  PubMed  CAS  Google Scholar 

  • Mahaffey DT, Moore MS, Brodsky FM, Anderson RGW (1989) Coat proteins isolated from clathrin coated vesicles can assemble into coated pits. J Cell Biol 108:1615–1624

    Article  PubMed  CAS  Google Scholar 

  • Marchisio PC, Cirillo D, Naldini L, Primavera MV, Teti A, Zambonin-Zallone A (1984) Cell-substratum interaction of cultured avian osteoclasts is mediated by specific adhesion structures. J Cell Biol 99:1696–1705

    Article  PubMed  CAS  Google Scholar 

  • Maupin P, Pollard TD (1983) Improved preservation and staining of HeLa cell actin filaments, clathrin-coated membranes, and other cytoplasmic structures by tannic acid-glutaraldehyde-saponin fixation. J Cell Biol 96:51–62

    Article  PubMed  CAS  Google Scholar 

  • Moore MS, Mahaffey DT, Brodsky FM, Anderson RGW (1987) Assembly of clathrin-coated pits onto purified plasma membranes. Science 236:558–563

    Article  PubMed  CAS  Google Scholar 

  • Nermut MV (1989) Strategy and tactics in electron microscopy of cell surfaces. Electron Microsc Rev 2:171–196

    Article  PubMed  CAS  Google Scholar 

  • Nermut MV, Eason P, Hirst EMA, Kellie S (1991) Cell/substratum adhesions in RSV-transformed rat fibroblasts. Exp Cell Res 193:382–397

    Article  PubMed  CAS  Google Scholar 

  • Nicol A, Nermut MV (1987) A new type of substratum adhesion structure in NRK cells revealed by correlated interference reflection and electron microscopy. Eur J Cell Biol 43:348–357

    PubMed  CAS  Google Scholar 

  • Nitsch L, Gionti E, Cancedda R, Marchisio PC (1989) The podosomes of Rous sarcoma virus transformed chondrocytes show a peculiar ultrastructural organization. Cell Biol Int Rep 13:919–926

    Article  PubMed  CAS  Google Scholar 

  • Ochoa GC, Slepnev VI, Neff L, Ringstad N, Takei K, Daniell L, Kim W, Cao H, McNiven M, Baron R, De Camilli P (2000)A functional link between dynamin and the actin cytoskeleton at podosomes.J Cell Biol 150:377–389

    Article  PubMed  CAS  Google Scholar 

  • Pumplin DW (1989) Acetylcholine receptor clusters of rat myotubes have at least three domains with distinctive cytoskeletal and membranous components. J Cell Biol 109:739–753

    Article  PubMed  CAS  Google Scholar 

  • Pumplin DW, Bloch RJ (1990) Clathrin-coated membrane: a distinct membrane domain in acetylcholine receptor clusters of rat myotubes. Cell Motil Cytoskelet 15:121–134

    Article  CAS  Google Scholar 

  • Robertson TA, Papadimitriou JM (1986) A morphometric study of coated pit formation on the subplasmalemmal surface of murine peritoneal macrophages after adhesion to glass. J Ultrastruct Mol Struct Res 96:125–135

    Article  PubMed  CAS  Google Scholar 

  • Saltel F, Destaing O, Bard F, Eichert D, Jurdic P (2004) Apatite-mediated actin dynamics in resorbing osteoclasts. Mol Biol Cell 15:5231–5241

    Article  PubMed  CAS  Google Scholar 

  • Samuelsson SJ, Luther PW, Pumplin DW, Bloch RJ (1993) Structures linking microfilament bundles to the membrane at focal contacts. J Cell Biol 122:485–496

    Article  PubMed  CAS  Google Scholar 

  • Samuelsson SJ, Luther PW, Bloch RJ (1996) Microfilament-membrane interactions in Xenopus myocytes. Cell Motil Cytoskelet 35:68–80

    Article  CAS  Google Scholar 

  • Spinardi L, Marchisio PC (2006) Podosomes as smart regulators of cellular adhesion. Eur J Cell Biol 85:191–194

    Article  PubMed  CAS  Google Scholar 

  • Svitkina TM, Verkhovsky AB, McQuade KM, Borisy GG (1997) Analysis of the actin-myosin II system in fish epidermal keratocytes: mechanism of cell body translocation. J Cell Biol 139:397–415

    Article  PubMed  CAS  Google Scholar 

  • Takemura R, Stenberg PE, Bainton DF, Werb Z (1986) Rapid redistribution of clathrin onto macrophage plasma membranes in response to Fc receptor-ligand interaction during frustrated phagocytosis. J Cell Biol 102:55–69

    Article  PubMed  CAS  Google Scholar 

  • Tarone G, Cirillo D, Giancotti FG, Comoglio PM, Marchisio PC (1985) Rous sarcoma virus-transformed fibroblasts adhere primarily at discrete protrusions of the ventral membrane called podosomes. Exp Cell Res 159:141–157

    Article  PubMed  CAS  Google Scholar 

  • Teti A, Marchisio PC, Zambonin-Zallone A (1991) Clear zone in osteoclast function: role of podosomes in regulation of bone-resorbing activity. Am J Physiol 261:C1–C7

    PubMed  CAS  Google Scholar 

  • Turksen K, Kanehisa J, Opas M, Heersche JNM, Aubin JE (1988) Adhesion patterns and cytoskeleton of rabbit osteoclasts on bone slices and glass. J Bone Miner Res 3:389–400

    PubMed  CAS  Google Scholar 

  • Vaananen HK, Horton M (1995) The osteoclast clear zone is a specialized cell-extracellular matrix adhesion structure. J Cell Sci 108:2729–2732

    PubMed  CAS  Google Scholar 

  • Verhovsky AB, Svitkina TM, Borisy GG (1997) Polarity sorting of actin filaments in cytochalasin-treated fibroblasts. J Cell Sci 110:1693–1704

    Google Scholar 

  • Vorobjev IA, Svitkina TM, Borisy GG (1997) Cytoplasmic assembly of microtubules in cultured cells. J Cell Sci 110:2635–2645

    PubMed  CAS  Google Scholar 

  • Woods A, Smith CG, Rees DA, Wilson G (1983)Stages in specialization of fibroblast adhesion and deposition of extracellular matrix.Eur J Cell Biol 32:108–116

    PubMed  CAS  Google Scholar 

  • Zambonin-Zallone A, Teti A, Grano M, Rubinacci A, Abbadini M, Gaboli M, Marchisio PC (1989) Immunocytochemical distribution of extracellular matrix receptors in human osteoclasts: a beta 3 integrin is colocalized with vinculin and talin in the podosomes of osteoclastoma giant cells. Exp Cell Res 182:645–652

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Oral Anatomy, Asahi University School of Dentistry, Hozumi 1851, Mizuho, Gifu, 501-0296, Japan

    Toshitaka Akisaka, Hisaho Yoshida & Reiko Suzuki

  2. Department of Oral Anatomy, Osaka Dental University, Hirakata 573-1121 Kuzuhahanazono 8-1, Osaka, Japan

    Keiko Takama

Authors
  1. Toshitaka Akisaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hisaho Yoshida
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Reiko Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Keiko Takama
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Toshitaka Akisaka.

Additional information

This work was supported by the following grants to T.A.: Grants-in-Aid for Scientific Research (C) (18592020) from the Ministry of Education, Science, and Culture of Japan and the Miyata Research Fund of Asahi University.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Akisaka, T., Yoshida, H., Suzuki, R. et al. Adhesion structures and their cytoskeleton-membrane interactions at podosomes of osteoclasts in culture. Cell Tissue Res 331, 625–641 (2008). https://doi.org/10.1007/s00441-007-0552-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-007-0552-x

Keywords

Search

Navigation