Skip to main content
SpringerLink
Log in

Immunocytochemical localization of vacuolar H+-ATPase and Cl−-HCO −3 anion exchanger (erythrocyte band-3 protein) in avian osteoclasts: Effect of calcium-deficient diet on polar expression of the H+-ATPase pump

  • Laboratory Investigations
  • Published:
Calcified Tissue International Aims and scope Submit manuscript

Abstract

Osteoclasts attach to the bone surface and resorb bone by secreting protons into an isolated subosteoclastic compartment. Previous studies have shown the presence of a vacuolar type H+-ATPase, and a functional Cl−-HCO −3 anion exchanger in the osteoclast. In the present studies, using a monoclonal antibody to the 31-kDa subunit of H+-ATPase and a rabbit antiserum to the erythrocyte band-3 protein (Cl−-HCO −3 anion exchanger) we have immunocytochemically localized the respective pumps in bone sections obtained from chickens fed a normal or a calcium-deficient diet for 4 weeks. Our results indicate that although H+-ATPase is either evenly distributed throughout the osteoclast or is more polarized at its ruffled membrane juxtaposed to the bone surface, the band-3 protein immunoreactivity is always localized to the plasma membrane which is not attached to the bone surface (basolateral membrane). Four weeks of a calcium-deficient diet resulted in a significant increase in the percentage of osteoclasts that were polarized for the H+-ATPase pump at their ruffled membrane, and a trend toward increased total number of osteoclasts, although the latter did not reach statistical significance (P =0.09). These changes were not accompanied by a significant increase in the intensity of staining for H+-ATPase. Band-3 protein immunoreactivity was always prominent, limited to the basolateral membrane, and did not alter with calcium-deficient diet or with changes in the degree of H+-ATPase polarization.

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

  1. Fallon MD (1984) Bone-resorbing fluid from osteoclasts is acidic. An in vitro micropuncture study. In: Cohn DV, Fujita T, Potts JT Jr, and Talmage RV (eds) Endocrine control of bone and calcium metabolism, Elsevier, Amsterdam, pp 144–146

    Google Scholar 

  2. Etherington DJ, Maciewicz RA, Taylor MAJ, Wardale RJ, Silver IA, Murrills RA, Pugh D (1986) The role of collagendegrading cysteine proteinases in connective tissue metabolism. In: Turk V (ed) Cysteine proteinases and their inhibitors. W. de Gruyter, Berlin, p 269

    Google Scholar 

  3. Anderson RE, Jee WSS, Woodbury DM (1985) Stimulation of carbonic anhydrase in osteoclasts by parathyroid hormone. Calcif Tissue Int 37:646–650

    PubMed  CAS  Google Scholar 

  4. Anderson RE, Woodbury DM, Jee WSS (1986) Humoral and ionic regulation of osteoclast acidity. Calcif Tissue Int 39: 252–258

    PubMed  CAS  Google Scholar 

  5. Baron R (1989) Polarity and membrane transport in osteoclasts. Connect Tissue Res 20:109–120

    PubMed  CAS  Google Scholar 

  6. Vaes G (1988) Cellular biology and biochemical mechanism of bone resorption. Clin Orthop 231:239–271

    PubMed  CAS  Google Scholar 

  7. Baron R, Neff L, Roy C, Bolsvert A, Caplan M (1986) Evidence for a high and specific concentration of Na+, K+-ATPase in the plasma membrane of the osteoclast. Cell 46: 311–320

    Article  PubMed  CAS  Google Scholar 

  8. Akisaka T, Gay CV (1986) An ultracytochemical investigation of ouabain-sensitive p-nitrophenylphosphatase in chick osteoclasts. Cell Tissue Res 244:57–62

    Article  PubMed  CAS  Google Scholar 

  9. Blair HC, Kahn AJ, Crouch EC, Jeffrey JJ, Teitelbaum SL (1986) Isolated osteoclasts resorb the organic and inorganic components of bone. J Cell Biol 102:1164–1172

    Article  PubMed  CAS  Google Scholar 

  10. Teitelbaum SL, Stewart CC, Kahn AJ (1979) Rodent peritoneal macrophages as bone-resorbing cells. Calcif Tissue Int 27:255–261

    Article  PubMed  CAS  Google Scholar 

  11. Brown D, Gluck S, Hartwig J (1987) Structure of the novel membrane-coating material in proton-secreting epithelial cells and identification as an H-ATPase. J Cell Biol 105:1637–1648

    Article  PubMed  CAS  Google Scholar 

  12. Balir HC, Teitelbaum SL, Ghiselli R, Gluck S (1989) Osteoclastic bone resorption by a polarized vacuolar proton pump. Science 245:855–857

    Article  Google Scholar 

  13. Vaananen HK, Karhukorpi EK, Sundquist K, Wallmark B, Roininen I, Hentunen T, Tuukkanen J, Lakkakorpi P (1990) Evidence for the presence of a proton pump of the vacuolar H+-ATPase type in the ruffled borders of osteoclasts. J Cell Biol 111:1305–1311

    Article  PubMed  CAS  Google Scholar 

  14. Akisaka T, Gay CV (1986) Ultracytochemical evidence for a proton-pump adenosine triphosphatase in chick osteoclasts. Cell Tissue Res 245:507–512

    Article  PubMed  CAS  Google Scholar 

  15. Teti A, Blair HC, Teitelbaum SL, Kahn AJ, Koziol C, Konsek J, Zambonin-Zallone A, Schlesinger PH (1989) Cytoplasmic pH regulation and chloride/bicarbonate exchange in avian osteoclasts, J Clin Invest 83:227–233

    PubMed  CAS  Google Scholar 

  16. Bastani B, Purcell H, Hemken Ph, Trigg D, Gluck S (1991) Expression and distribution of renal vacuolar protontranslocating adenosine triphosphatase in response to chronic acid and alkali loads in the rat. J Clin Invest 88:126–136

    PubMed  CAS  Google Scholar 

  17. Thomas HA, Machen TE, Smolka A, Baron R, Kopito RR (1989) Identification of a 185-kDa band-3-related polypeptide in oxyntic cells. Am J Physiol 257:C537-C544

    PubMed  CAS  Google Scholar 

  18. Baron R, Neff L, Louvard D, Courtoy PJ (1985) Cellmediated extracellular acidification and bone resorption: evidence for a low pH in resorbing lacunae and localization of a 100-kDa lysosomal membrane protein of the osteoclast ruffled border. J Cell Biol 101:2210–2222

    Article  PubMed  CAS  Google Scholar 

  19. Tuukkanen J, Vaananen HK (1986) Omeprazole, a specific inhibitor of H-K-ATPase, inhibits bone resorption in vitro. Calcif Tissue Int 38:123–125

    PubMed  CAS  Google Scholar 

  20. Kellokumpu S, Neff J, Jamsa-Kellokumpu S, Kopito R, Baron R (1988) A 115-kD polypeptide immunologically related to erythrocyte band-3 is present in golgi membranes. Science 242:1308–1311

    Article  PubMed  CAS  Google Scholar 

  21. Schuster VL, Fejes-Toth G, Naray-Fejes-Toth A, Gluck S (1991) Colocalization of H-ATPase and band 3 anion exchanger in rabbit-collecting duct intercalated cells. Am J Physiol 260:F506-F517

    PubMed  CAS  Google Scholar 

  22. Lorenzo JA, Raisz LG, Hock JM (1983) DNA synthesis is not necessary for osteoclastic responses to parathyroid hormone in cultured fetal rat long bones. J Clin Invest 72:1924–1929

    Article  PubMed  CAS  Google Scholar 

  23. Ibbotson KJ, Roodman GD, McManus LM, Mundy GR (1984) Identification and characterization of osteoclast-like cells and their progenitors in cultures of feline marrow mononuclear cells. J Cell Biol 99:471–480

    Article  PubMed  CAS  Google Scholar 

  24. Takahashi N, Mundy GR, Roodman GD (1986) Recombinant human interferon-α inhibits formation of human osteoclast-like cells. J Immunol 137:3544–3549

    PubMed  CAS  Google Scholar 

  25. MacDonald BR, Mundy GR, Clark S, Wang EA, Kuehl TJ, Stanley ER, Roodman GD (1986) Effects of human recombinant CSF-GM and highly purified CSF-1 on the formation of multinucleated cells with osteoclast characteristics in long-term bone marrow cultures. J Bone Miner Res 1:227–233

    Article  PubMed  CAS  Google Scholar 

  26. Schelling SH, Wolfe HJ, Tashjian AH (1980) Role of the osteoclast in prostaglandin E2-stimulated bone resorption. A correlative morphometric and biochemical analysis. Lab Invest 42:290–295

    PubMed  CAS  Google Scholar 

  27. Miller SC (1978) Rapid activation of the medullary bone osteoclast cell surface by parathyroid hormone. J Cell Biol 76: 615–618

    Article  PubMed  CAS  Google Scholar 

  28. Miller SC, Bowman BM, Myers RL (1984) Morphological and ultrastructural aspects of the activation of avian medullary bone osteoclasts by parathyroid hormone. Anat Rec 208:223–231

    Article  PubMed  CAS  Google Scholar 

  29. Holtrop ME, Raisz LG (1979) Comparison of the effects of 1,25-dihydroxycholecalciferol, prostaglandin E2 and osteoclast-activating factor with parathyroid hormone on the ultrastructure of osteoclasts in cultured long bones of fetal rats. Calcif Tissue Int 29:201–205

    Article  PubMed  CAS  Google Scholar 

  30. Holtrop ME, King GJ, Cox KA, Reit B (1979) Time-related changes in the ultrastructure of osteoclasts after injection of parathyroid hormone in young rats. Calcif Tissue Int 27:129–135

    Article  PubMed  CAS  Google Scholar 

  31. Miller SC, Jee WSS (1987) The bone lining cell: a distinct phenotype? Calcif Tissue Int 41:1–5

    PubMed  CAS  Google Scholar 

  32. Vanderwiel CJ (1980) An ultrastructural study of the components which make up the resting surface of bone. Metab Bone Dis Rel Res (suppl) 2:109

    Google Scholar 

  33. Zambonin-Zallone A, Teti A, Primavera MV (1984) Resorption of vital or devitalized bone by isolated osteoclasts in vitro. The role of lining cells. Cell Tissue Res 235:561–564

    Article  PubMed  CAS  Google Scholar 

  34. Malluche HH, Faugere MC (1986) Atlas of mineralized bone histology. Basel, Karger

    Google Scholar 

  35. Chambers TJ, Thomson BM, Fuller K (1984) Effects of substrate composition on bone resorption by rabbit osteoclasts. J Cell Sci 70:61–71

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Nephrology, St. Louis University Health Sciences Center, 3635 Vista Ave., 9N-FDT, 63110-0250, St. Louis, Missouri

    B. Bastani

  2. Department of Pathology, Jewish Hospital at Washington University, St. Louis, Missouri

    F. P. Ross

  3. Department of Biological Sciences, Stanford University, Stanford, California

    R. R. Kopito

  4. Washington University School of Medicine, St. Louis, Missouri

    S. L. Gluck

Authors
  1. B. Bastani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F. P. Ross
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R. R. Kopito
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S. L. Gluck
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Parts of this work have been presented in the American Society of Nephrology in November 1992

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bastani, B., Ross, F.P., Kopito, R.R. et al. Immunocytochemical localization of vacuolar H+-ATPase and Cl−-HCO −3 anion exchanger (erythrocyte band-3 protein) in avian osteoclasts: Effect of calcium-deficient diet on polar expression of the H+-ATPase pump. Calcif Tissue Int 58, 332–336 (1996). https://doi.org/10.1007/BF02509381

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02509381

Key words

Search

Navigation