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Carbonic anhydrase II gene transcript in cultured osteoclasts from neonatal rats: effect of calcitonin

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Abstract

Carbonic anhydrase II (CA II), an enzyme catalyzing the interconversion of CO2 and water to HCO 3 and protons, has a key role in osteoclastic bone resorption, but little is known of the regulation of CA II gene expression by calcitonin. Analysis of mRNA in osteoclasts has been difficult because of the problems of obtaining sufficient number of purified osteoclasts from bone. In this study, however, we have investigated the regulation of CA II mRNA in rat osteoclasts and their putative mononuclear precursors by using in situ hybridization. We have found that the CA II gene is expressed at high levels in osteoclasts and what are probably their maturing mononuclear precursors. Measurement of CA II mRNA in cultured osteoclasts and their putative mononuclear precursor cells by cytophotometry provided evidence that calcitonin, a direct inhibitor of mammalian osteoclast activity, reduces the levels of CA II mRNA in a dose dependent manner; maximum reduction was observed at a concentration of 100pM of calcitonin. In addition, calcitonin reduced the number of CA II mRNA-positive mononuclear precursor cells. The results also suggest that expression of the CA II gene is a feature of cells committed to the osteoclast lineage.

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Abbreviations

CA II :

Carbonic anhydrase II

TRAcP :

tartrate-resistant acid phosphatase

AcP :

acid phosphatase

MOPC :

mononuclear osteoclast precursor cells

References

  • Arnett TR, Dempster DW (1987) A comparative study of disaggregated chicken and rat osteoclasts in vitro: effect of calcitonin and prostaglandins. Endocrinology 120: 602–608

    Google Scholar 

  • Baron R (1989) Molecular mechanisms of bone resorption by osteoclasts. Anat Rec 224: 317–324

    Google Scholar 

  • Baron R, Neff L, Tran Van P, Nefussi JR, Vignery A (1986) Kinetic and cytochemical identification of osteoclast precusors and their differentiation into multinucleated osteoclasts. Am J Pathol 122: 363–378

    Google Scholar 

  • Billecocq A, Emanuel JR, Levenson R, Baron R (1990) 1α, 25-Dihydroxyvitamin D3 regulated the expression of carbonic anhydrase II in nonerythoid avaian bone marrow cells. Proc Natl Acad Sci USA 87: 6470–6474

    Google Scholar 

  • Blair HC, Teitlebaum SL, Ghiselli R, Glucks (1989) Osteoclastic bone resorption by a polarized vacuolar proton pump. Science 245: 855–857

    Google Scholar 

  • Brown D, Roth J, Kumpulainen T, Orci L (1982) Ultrastructural immunocytochemical localization of carbonic anhydrase. Histochemistry 75: 209–213

    Google Scholar 

  • Cathala G, Savouret JF, Mendez B, West BL, Karin M, Martial JA, Baxter JD (1983) A method for isolation of intact, translationally active ribonucleic acid. DNA Cell Biol 2: 329–335

    Google Scholar 

  • Chambers TJ, Revell PA, Fuller K, Athanasou NA (1984) Resorption of bone by isolated rabbit osteoclasts. J Cell Sci 66: 383–399

    Google Scholar 

  • Everts V, Delaisse JM, Korper W, Niehof A, Vaes G, Beertsen W (1992) Degradation of collagen in the bone-resorbing compartment underlying the osteoclast involve both cysteine-proteinases and matrix metalloproteinases. J Cell Physiol 150: 221–231

    Google Scholar 

  • Gay CV, Schraer H, Anderson RE, Cao H (1984) Current studies on the location and function of carbonic anhydrase in osteoclasts. Ann NY Acad Sci 429: 473–478

    Google Scholar 

  • Gluck KS, Roodman GD (1990) Sequential expression of phenotype markers for osteoclasts during differentiation of precursors for multinucleated cells formed in long term human marrow culture. Endocrinology 127: 3215–3221

    Google Scholar 

  • Hall TJ, Chambers TJ (1990) The Na+/H+ antiporter is the primary proton transport system used by osteoclasts during bone resorption. J Cell Physiol 142: 420–424

    Google Scholar 

  • Hall TJ, Higgins W, Tardif C, Chambers TJ (1991) A comparison of the effect of inhibitors of carbonic anhydrase on osteoclastic bone resorption and purified carbonic anhydrase isoenzyme II. Calcif Tissue Int 49: 328–332

    Google Scholar 

  • Marino LR, Muglia BH, Yamada T (1990) H+−K+-ATPase and carbonic anhydrase II gene expression in the developing rat fundus. Am J Physiol 258: G108-G115

    Google Scholar 

  • Muallem SD, Blissard EJ, Cragoo E, Sachs G (1988) Activation of the Na+/H+ and Cl/HCO 3 exchange by stimulation of acid secretion in the parietal cell. J Biol Chem 263: 14703–14711

    Google Scholar 

  • Nicholson GC, Moseley JM, Serton FA, Mendelsohn PAD, Martin TJ (1986) Abundant calcitonin receptors in isolated rat osteoclasts: biochemical and autoradiographic characterization. J Clin Invest 78: 355–360

    Google Scholar 

  • Nicholson GC, Moseley JM, Yate AFP, Martin TJ (1987) Control of cyclic adenosine 3′-5′-monophosphate production in osteoclasts: calcitonin-induced persistent activation and homologous desensitization of adenylate cyclase. Endocrinology 120: 1902–1908

    Google Scholar 

  • Overgaard K, Riis BJ, Christiansen C (1989) Nasal calcitonin for treatment of established osteoporosis. Clin Endocrinol (Oxf) 30: 435–442

    Google Scholar 

  • Prallet B, Male P, Neff L, Baron R (1992) Identification of a functional mononuclear precusor of the osteoclast in chicken medullary bone marrow cultures. J Bone Miner Res 7: 405–414

    Google Scholar 

  • Raisz LC, Simmons HA, Thompson WJ, Shepard KL, Anderson PS, Rodan GA (1988) Effect of a potent carbonic anhydrase inhibitor on bone resorption in organ culture. Endocrinology 122: 1083–1086

    Google Scholar 

  • Ries WL (1984) Osteogenic periosteum esterase activity: a comparative morphological and cytochemical study of bone cells in situ on rat proximal tibiae and in smears. J Histochem Cytochem 32: 55–62

    Google Scholar 

  • Roth DE, Venta PJ, Tashian RE, Sly WS (1992) Molecular basis of human carbonic anhydrase II deficiency. Proc Natl Acad Sci USA 89: 1804–1808

    Google Scholar 

  • Scheven BAA, Kawilarang-de Haas EWM, Wassenaar AM, Nijweide PJ (1986) Differentiation kinetics of osteoclasts in the periosteum of embryonic bones in vivo and in vitro. Anat Rec 214: 418–423

    Google Scholar 

  • Sundquist KT, Leppilampi M, Jarvelin K, Kumpulainen T, Vaananen HK (1987) Carbonic anhydrase isoenzyme in isolated rat peripheral monocytes, tissues macrophages and osteoclasts. Bone 8: 33–38

    Google Scholar 

  • Tashian RE, Hewett-Emmett D (1984) Biology and chemistry of the carbonic anhydrase. Ann NY Acad Sci 429: 1–64

    Google Scholar 

  • Vaananen HK (1984) Immunohistochemical localization of carbonic anhydrase isoenzymes I and II in human bone, cartilage and giant cell tumour. Histochemistry 81: 485–487

    Google Scholar 

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

    Google Scholar 

  • Zaidi M, Datta HK, Moonga BS, Maclntyre I (1990) Evidence that the action of calcitonin on rat osteoclasts is mediated by G proteins acting via separate post-receptor pathways. J Endocrinol 126: 473–481

    Google Scholar 

  • Zheng MH, Papadimitriou JM, Nicholson GC (1991) A quantitative cytochemical investigation of osteoclasts and multinucleate giant cells. Histochem J 23: 180–188

    Google Scholar 

  • Zheng MH, Papadimitriou JM, Nicholson GC (1991a) RNA synthesis in isolated rat ostoeclasts: inhibitory effect of calcitonin. Bone 12: 317–322

    Google Scholar 

  • Zheng MH, Nicholson GC, Warton A, Papadimitriou JM (1991b) What is new in osteoclast ontogeny? Pathol Res Pract 187: 117–125

    Google Scholar 

  • Zheng MH, Wood DJ, Papadimitriou JM, Nicholson GC (1992) Evidence that protein kinase-A, calcium-calmodulin kinase and cytoskeletal proteins are involved in osteoclast retraction induced by calcitonin. Exp Mol Pathol 57: 105–115

    Google Scholar 

  • Zheng MH, Fan Y, Wysocki S, Wood DJ, Papadimitriou JM (1993) Detection of mRNA for carbonic anhydrase II in human osteoclast-like cells by in-situ hybridization. J Bone Miner Res. 8: 111–116

    Google Scholar 

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Zheng, M.H., Fan, Y., Wysocki, S. et al. Carbonic anhydrase II gene transcript in cultured osteoclasts from neonatal rats: effect of calcitonin. Cell Tissue Res 276, 7–13 (1994). https://doi.org/10.1007/BF00354778

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  • DOI: https://doi.org/10.1007/BF00354778

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