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Avian medullary bone in organ culture: Effects of vitamin D metabolites on collagen synthesis

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Summary

A new organ culture system for the study of bone metabolism has been developed using chicken medullary bone. The presence of viable bone cells in culture was demonstrated by histological and histochemical techniques. Incorporation of3H-proline into collagenase-digestible protein (CDP) and noncollagen protein (NCP) was determined using purified bacterial collagenase. Collagen accounted for approximately 10–15% of the total protein labeled. The addition of 1,25-dihydroxycholecalciferol (1,25 (OH)2D3) resulted in a dose-dependent inhibition of3H-proline incorporation into CDP at doses from 10−10M to 10−7M, with maximal suppression reaching 30% of control. The effect was specific for collagen, since3H-proline incorporation into NCP was unaffected. Hydroxyproline analysis of bone explants and culture medium revealed a 1,25(OH)2D3-induced decrease in the3H-hydroxyproline content of the system (bone + medium), suggesting that the effect of 1,25(OH)2D3 is due to inhibition of collagen synthesis rather than enhanced collagen degradation, impaiored incorporation of collagen into bone matrix, or bone resorption Medullary bone collagen synthesis was not affected by 24,25(OH)2D3, either alone or in combination with 1,25(OH)2D3. Structure-activity studies of vitamin D metabolites showed that 1,25(OH)2D3 and 1,24,25(OH)3D3 were the most potent metabolites tested, followed by 1-alpha(OH)D3. 25(OH)D3 and 24,25(OH)2D3 had no effect at concentrations as high as 10−7M. These results indicate a possible role for vitamin D in the regulation of medullary bone formation during the reproductive cycle of the egg-laying hen, and suggest the potential utility of medullary bone as anin vitro model for the study of bone formation

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References

  1. Taylor TG (1965) Calcium-endocrine relationships in the laying hen. Proc Nutr Soc 24:49–54

    Article  PubMed  CAS  Google Scholar 

  2. Comar CL, Driggers JC (1949) Secretion of radioactive calcium in the hen's egg. Science 109:282

    Article  CAS  PubMed  Google Scholar 

  3. Mueller WJ, Schraer R, Schraer H (1964) Calcium metabolism and skeletal dynamics of laying pullets. J Nutr 84:20–26

    PubMed  CAS  Google Scholar 

  4. Kyes P, Potter TS (1934) Physiological marrow ossification in female pigeons. Anat Rec 60:377–379

    Article  Google Scholar 

  5. Bloom W, Bloom MA, McLean FC (1941) Calcification and ossification. Medullary bone changes in the reproductive cycle of female pigeons. Anat Rec 87:443–475

    Article  Google Scholar 

  6. Aszenzi A, Francois C, Bocciarelli DS (1963) On the bone induced by estrogens in birds. J Ultrastruct Res 8:491–505

    Article  Google Scholar 

  7. Simkiss K (1967) Calcium in reproductive physiology Chapman and Hall, Reinhold, London, New York

    Google Scholar 

  8. Fisher LW, Schraer H (1980) The glycosaminoglycans of estrogen-induced medullary bone in Japanese quail. Arch Biochem Biophys 205:396–403

    Article  PubMed  CAS  Google Scholar 

  9. Kusuhara S, Schraer H (1982) Cytology and autoradiography of estrogen-induced differentiation of avian endosteal cells. Calcif Tissue Int 34:352–358

    Article  PubMed  CAS  Google Scholar 

  10. Hunter SJ, Schraer H (1983) In vitro synthesis of proteoglycans associated with medullary bone in Japanese quail. Arch Biochem Biophys 220:272–279

    Article  PubMed  CAS  Google Scholar 

  11. Spanos E, Pike JW, Haussler MR, Colston KW, Evans IMA, Goldner AM, McCain TA, MacIntyre I (1976) Circulating I-α,25-dihydroxyvitamin D in the chicken: enhancement by injection of prolactin and during egg laying. Life Science 19:1751–1756

    Article  CAS  Google Scholar 

  12. Castillo L, Tanaka Y, Wineland MJ, Jowsey JO, DeLuca HF (1979) Production of 1,25-dihydroxyvitamin D3 and formation of medullary bone in the egg-laying hen. Endocrinology 104:1598–1606

    PubMed  CAS  Google Scholar 

  13. Abe E, Tanabe R, Suda T, Yoshiki S, Horikawa H, Masumura T, Sugahara M (1979) Circadian rhythm of 1-α,25-dihydroxyvitamin D3 production in egg-laying hens. Biochem Biophys Res Commun 88:500–507

    Article  PubMed  CAS  Google Scholar 

  14. Raisz LG, Trummel CL, Holick MF, DeLuca HF (1972) 1,25-dihydroxycholecalciferol, a potent stimulator of bone resorption in tissue culture. Science 175:768–769

    Article  PubMed  CAS  Google Scholar 

  15. Reynolds JJ, Holick MF, DeLuca HF (1973) The role of vitamin D metabolites in bone resorption. Calcif Tissue Res 12:295–310

    Article  PubMed  CAS  Google Scholar 

  16. Stern PH, Trummel CL, Schnoes HK, DeLuca HF (1975) Bone-resorbing activity of vitamin D metabolites and congeners in vitro: influence of hydrosyl substituents in the A ring. Endocrinology 97:1552–1558

    PubMed  CAS  Google Scholar 

  17. 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 

  18. Rosen V, Clark NB (1982) Effects of 25-hydroxyvitamin D3 and 1,25-dihydroxy vitamin D3 on embryonic chick bone in organ culture. J Exp Zool 224:97–101

    Article  PubMed  CAS  Google Scholar 

  19. Raisz LG, Mania DM, Gworek SC, Dietrich JW, Canalis EM (1978) Hormonal control of bone collagen synthesis in vitro: inhibitory effect of 1-hydroxylated vitamin D metabolites. Endocrinology 102:731–735

    PubMed  CAS  Google Scholar 

  20. Rowe DW, Kream BE (1982) Regulation of collagen synthesis in fetal rat calvaria by 1,25-dihydroxyvitamin D3. J Biol Chem 257:8009–8015

    PubMed  CAS  Google Scholar 

  21. Bringhurst FR, Potts Jr JT (1982) Effects of vitamin D metabolites and analogs on bone collagen synthesis in vitro Calcif Tissue Int 34:103–110

    Article  PubMed  CAS  Google Scholar 

  22. Hock JM, Kream BE, Raisz LG (1982) Autoradiographic study of the effect of 1,25-dihydroxyvitamin D3 on bone matrix synthesis in vitamin D-replete rats. Calcif Tissue Int 34:347–351

    Article  PubMed  CAS  Google Scholar 

  23. Kream BE, Jose M, Yamada S, DeLuca HF (1977) A specific high affinity-binding macromolecule for 1,25-dihydroxyvitamin D3 in fetal bone. Science 197:1086–1087

    Article  PubMed  CAS  Google Scholar 

  24. Chen TL, Hirst MA, Feldman D (1979) A receptor-like binding macromolecule for 1-α,25-dihydroxycholecalciferol in cultured mouse bone cells. J Biol Chem 254:7491–7494 7494

    PubMed  CAS  Google Scholar 

  25. Manolagas SC, Haussler MR, Deftos LJ (1981) 1,25-dihydroxyvitamin D3 receptor-like macromolecule in rat osteogenic sarcoma cell lines. J Biol Chem 255:4414–4417

    Google Scholar 

  26. Bingham PJ, Raisz LG (1974) Bone growth in organ culture: effects of phosphate and other nutrients on bone and cartilage. Calcif Tissue Res 14:31–48

    Article  PubMed  CAS  Google Scholar 

  27. Burstone MS (1962) Enzyme histochemistry. Academic Press, New York

    Google Scholar 

  28. Peterkofsky B, Diegelmann R (1971) Use of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins. Biochemistry 10:988–949

    Article  PubMed  CAS  Google Scholar 

  29. Dietrich JW, Canalis EM, Maina DM, Raisz LG (1976) Hormonal control of bone collagen synthesis in vitro: effects of parathyroid hormone and calcitonin. Endocrinology 98:943–949

    Article  PubMed  CAS  Google Scholar 

  30. Bienkowski RS (1984) Radiochemical purity of14C-proline: implications for measurement of intracellular collagen degradation. Collagen Rel Res 4:195–200

    CAS  Google Scholar 

  31. Snedecor GW, Cochran WG (1980) Statistical methods. Iowa State University Press, Ames, Iowa

    Google Scholar 

  32. Canalis E (1983) Effect of hormones and growth factors on alkaline phosphatase activity and collagen synthesis in fetal rat calvariae. Metabolism 32:14–20

    Article  PubMed  CAS  Google Scholar 

  33. Rodan GA, Martin TJ (1981) Role of osteolasts in hormonal control of bone resorption—a hypothesis. Calcif Tissue Int 33:349–351

    Article  PubMed  CAS  Google Scholar 

  34. Wong GL (1984) Paracrine interactions in bone-secreted products of osteoblasts permit osteoclasts to respond to parathyroid hormone. J Biol Chem 259:4019–4022

    PubMed  CAS  Google Scholar 

  35. Genovese C, Rowe DW, Kream BE (1984) Construction of DNA sequences complementary to rat α1 and α2 collagen mRNA and their use in studying the regulation of type I collagen synthesis by 1,25-dihydroxyvitamin D. Biochemistry 23:6210–6216

    Article  PubMed  CAS  Google Scholar 

  36. Bienkowski RS (1983) Intracellular degradation of newly synthesized secretory proteins. Biochem J 214:1–10

    PubMed  CAS  Google Scholar 

  37. Kream BE, Rowe DW, Gworek SC, Raisz LG (1980) Parathyroid hormone alters collagen synthesis and procollagen mRNA levels in fetal rat calvaria. Proc Natl Acad Sci USA 77:5654–5658

    Article  PubMed  CAS  Google Scholar 

  38. Raisz LG, Kream BE, Smith MD, Simmons HA (1980) Comparison of the effects of vitamin D metabolites on collagen synthesis and resorption of fetal rat bone in organ culture. Calcif Tissue Int 32: 135–138

    Article  PubMed  CAS  Google Scholar 

  39. Ornoy A, Goodwin D, Noff D, Edelstein S (1978) 24,25-dihydroxyvitamin D is a metabolite of vitamin D essential for bone formation. Nature 276:517–519

    Article  PubMed  CAS  Google Scholar 

  40. Malluche HH, Henry H, Meyer-Sabellek W, Sherman D, Massry SG, Norman AW (1980) Effects and interactions of 24R,25(OH)2D3 and 1,25(OH)2D3 on bone. Am J Physiol 238: E494-E498

    PubMed  CAS  Google Scholar 

  41. Hurwitz S, Bar A, Cohen I (1973) Regulation of calcium absorption by fowl intestine. Am J Physiol 225:150–154

    PubMed  CAS  Google Scholar 

  42. Takahashi N, Shinki T, Abe E, Horiuchi N, Yamaguchi A Yoshiki S, Suda T (1983) The role of vitamin D in the medullary bone formation in egg-laying Japanese quail and in immature chicks treated with sex hormones. Calcif Tissue Int 35:465–471

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Physiology and Neurobiology, The University of Connecticut, 06268, Storrs, CT

    John R. Harrison & Nancy B. Clark

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  1. John R. Harrison
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  2. Nancy B. Clark
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Harrison, J.R., Clark, N.B. Avian medullary bone in organ culture: Effects of vitamin D metabolites on collagen synthesis. Calcif Tissue Int 39, 35–43 (1986). https://doi.org/10.1007/BF02555738

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  • DOI: https://doi.org/10.1007/BF02555738

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