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Ca^<2+> kinetics in mature osteoclast

Research Project

Project/Area Number 16591847
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Morphological basic dentistry
Research InstitutionMatsumoto Dental University

Principal Investigator

YAMAKI Mariko  Matsumoto Dental University, Oral Science, Hard Tissue Research, Assistant Professor, 総合歯科医学研究所, 講師 (90360221)

Co-Investigator(Kenkyū-buntansha) OZAWA Hidehiro  Graduate School of Oral Science, Hard Tissue Research, Professor, 大学院・歯学独立研究科, 教授 (60018413)
TAKAHASHI Naoyuki  Graduate School of Oral Science, Hard Tissue Research, Professor, 大学院・歯学独立研究科, 教授 (90119222)
SAHARA Noriyuki  Graduate School of Oral Science, Hard Tissue Research, Professor, 大学院・歯学独立研究科, 教授 (70064699)
HOSOYA Akihiro  School of Dentistry, Assistant, 歯学部, 助手 (70350824)
奥村 茂樹  松本歯科大学, 歯学部, 助手 (80350825)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥2,400,000 (Direct Cost: ¥2,400,000)
KeywordsOsteoclast / Ca^<2+> / Bone matrix / Endocytosis / Transportation in cell / Transcytosis / Vacuole / Lysosome / Ca^<+2>
Research Abstract

Osteoclasts play an important role in the metabolism of both organic and inorganic components in bone via the following steps-osteoclasts originate from bone marrow cells, differentiate with the aid of osteoblasts, fuse to each other and form mature, multi-nuclear gigantic cells, attach onto a bone surface with many podosomes (i.e. fibrous actin (F-actin^1) and integrin complexes), form actin rings, develop projecting structures (called ruffled borders), excrete substances including acids and proteolytic enzymes from the ruffled borders, solubilize bone, and leave resorption lacunae on the bone surface. Actin rings and ruffled borders are the morphological hallmarks of mature osteoclasts, and are therefore routinely used to identify mature osteoclasts. Osteoclasts undergo ‘transcytosis' of organic components, such as collagen, that are thus solubilized from bone via the following steps-the cells absorb the organic components via endocytosis at the ruffled borders, in-cell transport the … More se components to the apical membrane side of the cell, and perhaps expel (i.e.evacuate) them from the apical membrane toward the extracellular space. Thus, osteoclasts play a central role throughout the metabolism of organic components in bone-not only in the solubilization from bone but also in transcytosis steps (i.e. endocytosis at the ruffled borders, in-cell transportation, and evacuation from the apical membrane toward the extracellular). Although the role of osteoclasts in the metabolism of organic components is well documented, knowledge of the role of osteoclasts in the entire process of metabolism of Ca in bone has been fragmentary because no one has succeeded in directly observing such Ca in osteoclasts. Blood vessels actively absorb Ca released from bone by osteoclasts. Three hypothetical pathways for this uptake of Ca have been propose : (i)directly from resorption lacunae (i.e.the direct resorption pathway) ; (ii)through the intermediary of osteoclasts via Ca-specific channels (i.e.the channel-mediated transport pathway) ; (iii)through the intermediary of osteoclasts via transcytosis (i.e.the transcytosis transport pathway), which is analogous to the pathway for organic components. Although there is yet no evidence that definitively identifies any of these pathways or that refutes the transcytosis pathway, many researchers currently believe that the direct pathway is the main mechanism and that the channel-mediated pathway is a secondary mechanism. Among the three pathways, the transcytosis pathway seems to be least favored, only because no one has succeeded in directly observing Ca originating from bone in osteoclasts. To identify if the transcytosis pathway exists in osteoclasts, here we developed a method uses either fixed or living osteoclast-like cells (OCL) that were previously differentiated in vitro, uses a Ca-specific fluorescent dye (Fura Red^<TM>), to which a cell membrane is impermeable, and uses confocal laser scanning microscopy (LSM). We call this method the cell-membrane-impermeable dye (CMID) method. We used OCL because they are considered a good model of in vivo osteoclasts. Because the dye Fura Red^<TM> does not permeate an OCL membrane, the dye that exists inside OCL indicates active uptake, probably endocytosis, of the dye by OCL. If the Ca co-exists with such a dye inside OCL, direct and specific visualization of this Ca should be possible by detecting the emission from a dye-Ca complex. The Ca originating from bone might permeate OCL via two mechanisms : endocytosis and channels. If the quantity of such Ca is small (i.e.Ca via channels), the Ca would not be detectable, whereas if the quantity is large (i.e.Ca via endocytosis), it would be readily detectable and thus visualized. We therefore expected direct and specific visualization of the Ca via endocytosis by the CMID method. By using this method, we were successful in obtaining the first direct evidence that suggests massive Ca uptake occurs via endocytosis and that the Ca is transported toward the apical membrane side in OCL. This evidence also suggests that osteoclasts play an important role throughout the metabolism of Ca in bone. Less

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (21 results)

All 2006 2005 2004

All Journal Article (20 results) Book (1 results)

  • [Journal Article] Coordination of microtubules and the actin cytoskeleton is important in osteoclast function, but Calcitonin disrupts sealing zones without affecting microtubules networks.2006

    • Author(s)
      Okumura S.
    • Journal Title

      Bone (in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Transcytosis of calcium from bone by osteoclast-like cells evidenced by direct visualization of calcium in cells.2005

    • Author(s)
      Yamaki M.
    • Journal Title

      Arch. Biochem. Biophys. 440

      Pages: 10-17

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Effects of fixation and decalcification on the immunohistochemical localization of bone matrix proteins in fresh-frozen bone sections.2005

    • Author(s)
      Hosoya A.
    • Journal Title

      Histochem Cell Biol 123・6

      Pages: 639-646

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Prostaglandin E2 Receptors EP2 and EP4 Are Down-regulated during Differentiation of Mous Osteoclasts from Their Precursors.2005

    • Author(s)
      Kobayashi Y.
    • Journal Title

      J. Biol. Chem. 280・25

      Pages: 24035-24042

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Prostaglandin E2 enhances osteoclastic differentiation of precursor cells through protein kinase A-dependent phosphorylation of TAKI2005

    • Author(s)
      Kobayashi Y.
    • Journal Title

      J. Biol. Chem. 280・12

      Pages: 11395-11403

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Distribution of actin filaments, non-muscle myosin, M-Ras, and extracellular signal-regulated kinase (ERK) in osteoclasts after calcitionin administration.2005

    • Author(s)
      Nakamura H.
    • Journal Title

      Arch. Histol. Cytol. 68・2

      Pages: 143-150

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Transcytosis of calcium from bone by osteoclast-like cells evidenced by direct visualization of calcium in cells.2005

    • Author(s)
      Yamaki M.
    • Journal Title

      Arch.Biochem.Biophys. 440

      Pages: 10-17

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Effects of fixation and decalcification on the immunohistochemical localization of bone matrix proteins in fresh-frozen bone sections.2005

    • Author(s)
      Hosoya A.
    • Journal Title

      Histochem.Cell Biol. 123・6

      Pages: 639-646

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Prostaglandin E2 Receptors EP2 and EP4 Are Down-regulated during Differentiation of Mous Osteoclasts from Their Precursors.2005

    • Author(s)
      Kobayashi Y.
    • Journal Title

      J.Biol.Chem. 280■25

      Pages: 24035-24042

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Prostaglandin E2 enhances osteoclastic differentiation of precursor cells through protein kinase A-dependent phosphorylation of TAK1.2005

    • Author(s)
      Kobayashi Y.
    • Journal Title

      J.Biol.Chem. 280■12

      Pages: 11395-11403

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Distribution of actin filaments, non-muscle myosin, M-Ras, and extracellular signal-regulated kinase(ERK) in osteoclasts after calcitionin administration.2005

    • Author(s)
      Nakamura H.
    • Journal Title

      Arch.Histol.Cytol. 68・2

      Pages: 143-150

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Prostaglandin E2 Receptors EP2 and EP4 Are Down-regulated during Differentiation of Mous Osteoclasts from Their Precursors.2005

    • Author(s)
      Kobayashi Y.
    • Journal Title

      J.Biol.Chem. 280・25

      Pages: 24035-24042

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Prostaglandin E2 enhances osteoclastic differentiation of precursor cells through protein kinase A-dependent phosphorylation of TAK1.2005

    • Author(s)
      Kobayashi Y.
    • Journal Title

      J.Biol.Chem. 280・12

      Pages: 11395-11403

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Prostaglandin E_2 strongly inhibits human osteoclast formation2005

    • Author(s)
      Take I.
    • Journal Title

      Endocrinology 146・12

      Pages: 5204-5214

    • Related Report
      2005 Annual Research Report
  • [Journal Article] MyoD88 But Not TRIF Is Essential for Osteoclastogenesis Induced by Lipop olysaccharide, Diacyl Lipopeptide, and IL-1α.2004

    • Author(s)
      Sato N
    • Journal Title

      J.Exp.Med. 200.5

      Pages: 601-611

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Cementum-Like Tissue Deposition on the Resorbed Enamel Surface of Human Deciduous Teeth Prior to Shedding.2004

    • Author(s)
      Sahara N
    • Journal Title

      Anat Rec PART A 279.2

      Pages: 779-791

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Periodontal regeneration of transplanted rat molars after cryopreservation.2004

    • Author(s)
      Kawasaki N
    • Journal Title

      Arch Oral Biol 49.1

      Pages: 59-69

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Calcitonin-induced change in serum calcium levels and its relationship to osteoclast morphology and number of calcitonin receptors.2004

    • Author(s)
      Ikegame M
    • Journal Title

      BONE 35.1

      Pages: 27-33

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Histochemical evaluation for the biological effect of menatetrenone on metaphyseal trabeculae of ovariectomized rats.2004

    • Author(s)
      Asawa Y
    • Journal Title

      BONE 35.4

      Pages: 870-880

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Osteocytic osteolysis observed in rats to which parathyroid hormone was continuously administered.2004

    • Author(s)
      Tazawa K
    • Journal Title

      J Bone Miner Metab 22.6

      Pages: 524-529

    • Related Report
      2004 Annual Research Report
  • [Book] Hard Tissue 硬組織研究ハンドブック2005

    • Author(s)
      松本歯科大学大学院 硬組織研究グループ
    • Total Pages
      261
    • Publisher
      MDU出版会
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary

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Published: 2004-04-01   Modified: 2016-04-21  

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