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Modulation of osteoclastogenesis through adrenomedullin receptors on osteoclast precursors: initiation of differentiation by asymmetric cell division


Adrenomedullin (ADM), a member of the calcitonin family of peptides, is a potent vasodilator and was shown to have the ability to modulate bone metabolism. We have previously found a unique cell surface antigen (Kat1 antigen) expressed in rat osteoclasts, which is involved in the functional regulation of the calcitonin receptor (CTR). Cross-linking of cell surface Kat1 antigen with anti-Kat1 antigen monoclonal antibody (mAbKat1) stimulated osteoclast formation only under conditions suppressed by calcitonin. Here, we found that ADM provoked a significant stimulation in osteoclastogenesis only in the presence of calcitonin; a similar biological effect was seen with mAbKat1 in the bone marrow culture system. This stimulatory effect on osteoclastogenesis mediated by ADM was abolished by the addition of mAbKat1. 125I-labeled rat ADM (125I-ADM)-binding experiments involving micro-autoradiographic studies demonstrated that mononuclear precursors of osteoclasts abundantly expressed ADM receptors, and the specific binding of 125I-ADM was markedly inhibited by the addition of mAbKat1, suggesting a close relationship between the Kat1 antigen and the functional ADM receptors expressed on cells in the osteoclast lineage. ADM receptors were also detected in the osteoclast progenitor cells in the late mitotic phase, in which only one daughter cell of the dividing cell express ADM receptors, suggesting the semiconservative cell division of the osteoclast progenitors in the initiation of osteoclastogenesis. Messenger RNAs for the receptor activity-modifying-protein 1 (RAMP1) and calcitonin receptor-like receptor (CRLR) were expressed in cells in the osteoclast lineage; however, the expression of RAMP2 or RAMP3 was not detected in these cells. It is suggested that the Kat1 antigen is involved in the functional ADM receptor distinct from the general ADM receptor, consisting of CRLR and RAMP2 or RAMP3. Modulation of osteoclastogenesis through functional ADM receptors abundantly expressed on mononuclear osteoclast precursors is supposed to be important in the fine regulation of osteoclast differentiation in a specific osteotrophic hormonal condition with a high level of calcitonin in blood.

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Fig. 1: Histological relationship between osteoclasts and blood vessels.
Fig. 2: Stimulation of osteoclastic MNC formation by ADM only when cells are treated with calcitonin.
Fig. 3: Osteoclast-specific mAbKat1 blocked ADM-induced osteoclastogenesis in the presence of calcitonin.
Fig. 4: Detection of specific receptors for ADM on cells in culture of osteoclastogenesis and inhibition of 125I-ADM binding to cells in the osteoclast lineage by mAbKat1.
Fig. 5: Detection of specific receptors for ADM on osteoclastic cell lineage analyzed by autoradiography and RT-PCR.
Fig. 6: Schematic demonstration of the expression of functional ADM receptors related to osteoclast-specific Kat1 antigen in cells in the osteoclast lineage.


  1. 1.

    Kitamura K, Kangawa K, Kawamoto M, Ichiki Y, Nakamura S, Matsuo H, et al. Adrenomedullin: a novel hypotensive peptide isolated from human pheochromocytoma. Biochem Biophys Res Commun. 1993;192:553–60.

    CAS  Article  Google Scholar 

  2. 2.

    Xie J, Guo J, Kanwal Z, Wu M, Lv X, Ibrahim NA, et al. Calcitonin and bone physiology: in vitro, in vivo, and clinical investigations. Int J Endocrinol. 2020;2020:3236828.

    PubMed  PubMed Central  Google Scholar 

  3. 3.

    McLatchie LM, Fraser NJ, Main MJ, Wise A, Brown J, Tompson N, et al. RAMPs regulate the transport and ligand specificity of the calcitonin-receptor-like receptor. Nature. 1998;393:333–39.

    CAS  Article  Google Scholar 

  4. 4.

    Fischer JP, Els-Heindl S, Beck-Sickinger A. Adrenomedullin—current perspective on a peptide hormone with significant therapeutic potential. Peptides. 2020;131:170347.

    CAS  Article  Google Scholar 

  5. 5.

    Geven C, Kox M, Pickkers P. Adrenomedullin and adrenomedullin-targeted therapy as treatment strategies relevant for sepsis. Front Immunol. 2018;9:292

    Article  Google Scholar 

  6. 6.

    Hippenstiel S, Witzenrath M, Schmeck B, Hocke A, Krisp M, Krull M, et al. Adrenomedullin reduces endothelial hyperpermeability. Circ Res. 2002;91:618–25.

    CAS  Article  Google Scholar 

  7. 7.

    Langen UH, Pitulescu ME, Kim JM, Enriquez-Gasca R, Sivaraj KK, Kusambe AP, et al. Cell-matrix signals specify bone endothelial cells during developmental osteogenesis. Nat Cell Biol. 2017;19:189–201.

    CAS  Article  Google Scholar 

  8. 8.

    Ramasamy SK, Kusumbe AP, Wang L, Adams RH. Endothelial Notch activity promotes angiogenesis and osteogenesis in bone. Nature. 2014;507:376–80.

    CAS  Article  Google Scholar 

  9. 9.

    Kusumbe AP, Ramasamy SK, Adams RH. Coupling of angiogenesis and osteogenesis by a specific vessel subtype in bone. Nature. 2014;507:323–28.

    CAS  Article  Google Scholar 

  10. 10.

    Yasuda H, Shima N, Nakagawa N, Yamaguchi K, Kinosaki M, Mochizuki S, et al. Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci USA. 1998;95:3597–602.

    CAS  Article  Google Scholar 

  11. 11.

    Lacey DL, Timms E, Tan HL, Kelley MJ, Dunstan CR, Burgess T, et al. Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell. 1998;93:165–76.

    CAS  Article  Google Scholar 

  12. 12.

    Shiratori T, Kyumoto-Nakamura Y, Kukita A, Uehara N, Zhang JQ, Koda K, et al. IL-1beta Induces pathologically activated osteoclasts bearing extremely high levels of resorbing activity: a possible pathological subpopulation of osteoclasts, accompanied by suppressed expression of kindlin-3 and talin-1. J Immunol. 2018;200:218–28.

    CAS  Article  Google Scholar 

  13. 13.

    Xie H, Cui Z, Wang L, Xia Z, Hu Y, Xian L, et al. PDGF-BB secreted by preosteoclasts induces angiogenesis during coupling with osteogenesis. Nat Med. 2014;20:1270–78.

    CAS  Article  Google Scholar 

  14. 14.

    Cappariello A, Maurizi A, Veeriah V, Teti A. The Great Beauty of the osteoclast. Arch Biochem Biophys. 2014;558:70–78.

    CAS  Article  Google Scholar 

  15. 15.

    Charles JF, Aliprantis AO. Osteoclasts: more than ‘bone eaters’. Trends Mol Med. 2014;20:449–59.

    CAS  Article  Google Scholar 

  16. 16.

    Kukita T, Kukita A, Nagata K, Maeda H, Kurisu K, Watanabe T, et al. Novel cell-surface Ag expressed on rat osteoclasts regulating the function of the calcitonin receptor. J Immunol. 1994;153:5265–73.

    CAS  PubMed  Google Scholar 

  17. 17.

    Kukita T, Kukita A, Xu L, Maeda H, Iijima T. Successful detection of active osteoclasts in situ by systemic administration of an osteoclast-specific monoclonal antibody. Calcif Tissue Int. 1998;63:148–53.

    CAS  Article  Google Scholar 

  18. 18.

    Kukita T, Kukita A, Xu L, Watanabe T, Iijima T. Kat1-antigen–a reliable immunological marker for identifying osteoclast precursors of rats: detection of subpopulations among precursors and initiation of osteoclastogenesis. Histochem Cell Biol. 2001;115:215–22.

    CAS  Article  Google Scholar 

  19. 19.

    Matsubara R, Kukita T, Ichigi Y, Takigawa I, Qu PF, Funakubo N, et al. Characterization and identification of subpopulations of mononuclear preosteoclasts induced by TNF-alpha in combination with TGF-beta in rats. PLoS ONE. 2012;7:e47930.

    CAS  Article  Google Scholar 

  20. 20.

    Langston AL, Ralston SH. Management of Paget’s disease of bone. Rheumatology 2004;43:955–59.

    CAS  Article  Google Scholar 

  21. 21.

    Kukita T, Kukita A, Watanabe T, Iijima T. Osteoclast differentiation antigen, distinct from receptor activator of nuclear factor kappa B, is involved in osteoclastogenesis under calcitonin-regulated conditions. J Endocrinol. 2001;170:175–83.

    CAS  Article  Google Scholar 

  22. 22.

    Kuratani T, Nagata K, Kukita T, Hotokebuchi T, Nakasima A, Iijima T, et al. Induction of abundant osteoclast-like multinucleated giant cells in adjuvant arthritic rats with accompanying disordered high bone turnover. Histol Histopathol. 1998;13:751–59.

    CAS  PubMed  Google Scholar 

  23. 23.

    Li YJ, Kukita A, Teramachi J, Nagata K, Wu Z, Akamine A, et al. A possible suppressive role of galectin-3 in upregulated osteoclastogenesis accompanying adjuvant-induced arthritis in rats. Lab Invest. 2009;89:26–37.

    CAS  Article  Google Scholar 

  24. 24.

    Moriyama K, Kukita A, Li YJ, Uehara N, Zhang JQ, Takahashi I, et al. Regulation of osteoclastogenesis through Tim-3: possible involvement of the Tim-3/galectin-9 system in the modulation of inflammatory bone destruction. Lab Invest. 2014;94:1200–11.

    CAS  Article  Google Scholar 

  25. 25.

    Takahashi N, Yamana H, Yoshiki S, Roodman GD, Mundy GR, Jones SJ, et al. Osteoclast-like cell formation and its regulation by osteotropic hormones in mouse bone marrow cultures. Endocrinology. 1988;122:1373–82.

    CAS  Article  Google Scholar 

  26. 26.

    Kukita A, Kukita T, Shin JH, Kohashi O. Induction of mononuclear precursor cells with osteoclastic phenotypes in a rat bone marrow culture system depleted of stromal cells. Biochem Biophys Res Commun. 1993;196:1383–89.

    CAS  Article  Google Scholar 

  27. 27.

    Badawy T, Kyumoto-Nakamura Y, Uehara N, Zhang J, Sonoda S, Hiura H, et al. Osteoblast lineage-specific cell-surface antigen (A7) regulates osteoclast recruitment and calcification during bone remodeling. Lab Invest. 2019;99:866–84.

    CAS  Article  Google Scholar 

  28. 28.

    Romeo SG, Alawi KM, Rodrigues J, Singh A, Kusumbe AP, Ramasamy SK. Endothelial proteolytic activity and interaction with non-resorbing osteoclasts mediate bone elongation. Nat Cell Biol. 2019;21:430–41.

    CAS  Article  Google Scholar 

  29. 29.

    Zaidi M, Alam AS, Bax BE, Shankar VS, Bax CMR, Gill JS, et al. Role of the endothelial cell in osteoclast control: new perspectives. Bone. 1993;14:97–102.

    CAS  Article  Google Scholar 

  30. 30.

    Ibrahimi Disha S, Furlani B, Drevensek G, Plut A, Yanagisawa M, Hudoklin S, et al. The role of endothelin B receptor in bone modelling during orthodontic tooth movement: a study on ETB knockout rats. Sci Rep. 2020;10:14226.

    CAS  Article  Google Scholar 

  31. 31.

    Kukita T, Hata K, Kukita A, Iijima T. Laminin, a major basement membrane component of the blood vessel, as a negative regulator of osteoclastogenesis. Calcif Tissue Int. 1998;63:140–42.

    CAS  Article  Google Scholar 

  32. 32.

    Uehara N, Kukita A, Kyumoto-Nakamura Y, Yamaza T, Yasuda H, Kukita T. Osteoblast-derived Laminin-332 is a novel negative regulator of osteoclastogenesis in bone microenvironments. Lab Invest. 2017;97:1235–44.

    CAS  Article  Google Scholar 

  33. 33.

    Naot D, Callon KE, Grey A, Cooper JS, Reid IR, Cornish J. A potential role for adrenomedullin as a local regulator of bone growth. Endocrinology. 2001;142:1849–57.

    CAS  Article  Google Scholar 

  34. 34.

    Cornish J, Naot D, Reid IR. Adrenomedullin—a regulator of bone formation. Regul Pept. 2003;112:79–86.

    CAS  Article  Google Scholar 

  35. 35.

    Liu Y, Zuo G, Meng X, Gao X, Zhang L, Tang P. Adrenomedullin inhibits osteoclast differentiation through the suppression of receptor activator of nuclear factor-kappaB ligand-induced nuclear factor-kappaB activation in glucocorticoid-induced osteoporosis. Exp Ther Med. 2017;14:4009–16.

    CAS  PubMed  PubMed Central  Google Scholar 

  36. 36.

    Katajisto P, Dohla J, Chaffer CL, Pentinmikko N, Marjanovie N, Iqbal S, et al. Stem cells. Asymmetric apportioning of aged mitochondria between daughter cells is required for stemness. Science. 2015;348:340–43.

    CAS  Article  Google Scholar 

  37. 37.

    Inaba M, Buszczak M, Yamashita YM. Nanotubes mediate niche-stem-cell signalling in the Drosophila testis. Nature. 2015;523:329–32.

    CAS  Article  Google Scholar 

  38. 38.

    Takahashi S, Goldring S, Katz M, Hilsenbeck S, Williams R, Roodman GD. Downregulation of calcitonin receptor mRNA expression by calcitonin during human osteoclast-like cell differentiation. J Clin Invest. 1995;95:167–71.

    CAS  Article  Google Scholar 

  39. 39.

    Krieger NS, Feldman RS, Tashjian AH. Parathyroid hormone and calcitonin interactions in bone: irradiation-induced inhibition of escape in vitro. Calcif Tissue Int. 1982;34:197–203.

    CAS  Article  Google Scholar 

  40. 40.

    Nakamura T, Toyofuku F, Kanda S. Whole-body irradiation inhibits the escape phenomenon of osteoclasts in bones of calcitonin-treated rats. Calcif Tissue Int. 1985;37:42–45.

    CAS  Article  Google Scholar 

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Author information




TK and AK performed the study concept and design. TK, HH, TY, IT, and AK performed the development of methodology and writing, review, and revision of the paper. TK, HH, and AK provided acquisition of data and statistical analysis. JG, JZ, YK-N, NU, SM, and SS provided technical and material support. All authors read and approved the final paper.

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Correspondence to Toshio Kukita.

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This work was supported in part by a Grant for Scientific Research from the Japanese Ministry of Education, Science and Culture (Project 09671858, 15K15677, and 18K09506).

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The authors declare no competing interests.

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Kukita, T., Hiura, H., Gu, JY. et al. Modulation of osteoclastogenesis through adrenomedullin receptors on osteoclast precursors: initiation of differentiation by asymmetric cell division. Lab Invest 101, 1449–1457 (2021).

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