Summary
The effect of parathyroid hormone (PTH) on alkaline phosphatase activity was examined in confluent, serum-free primary cultures of neonatal mouse calvarial cells. It was found that synthetic bPTH-(1-34) caused an increase in the specific activity of skeletal alkaline phosphatase isoenzyme by 18 hours. Between 10 and 500 ng/ml, the mganitude of the change was directly related to peptide concentration. The change occurred in the absence of any effect on cell number, total cell protein, or DNA and was not the result of an effect on either proliferation or survival of a specific cell population. Results of histochemical studies indicate that bPTH-(1-34) caused an increase in the proportion of cells containing enzyme activity. The response was duplicated by intact bPTH-(1-84) and DBcAMP, but not by oxidized bPTH-(1-34) or insulin and did not require prostaglandin synthesis or hydroxylation of 25-hydroxyvitamin D3. These results demonstrate that bPTH has a direct effect on osteoblast maturationin vitro, that the effect is specific for PTH, and suggest that it is mediated by cAMP.
Similar content being viewed by others
References
Young RW (1964) Specialization of bone cells. In: Frost HM (ed) Bone biodynamics. Charles C Thomas, Springfield, Ill, p 117
McGuire JL, Marks SC Jr (1974) The effects of parathyroid hormone on bone cell structure and function. Clin Orthop Rel Res 100:392–405
Gaillard PJ (1961) Parathyroid and bone in tissue culture. In: Greep RO, Talmage RV (eds) The parathyroids. Charles C Thomas Springfield, Ill, p 20
Raisz LG (1965) Bone resorption in tissue culture. Factors influencing the response to parathyroid hormone. J Clin Invest 44:103–116
Goldhaber P (1966) Remodeling of bone in tissue culture. J Dent Res 45:490–499
Dietrich JW, Canalis EM, Maina DM, Raisz LG (1976) Hormonal control of bone collagen synthesisin vitro: Effects of parathyroid hormone and calcitonin. Endocrinology 98:943–949
Rosen DM, Luben RA (1983) Multiple hormonal mechanisms for the control of collagen synthesis in an osteoblast-like cell line, MMB-1. Endocrinology 112:992–999
Howard GA, Bottemiller BL, Baylink DJ (1980) Evidence for the coupling of bone formation to bone resorptionin vitro. Metab Bone Dis Rel Res 2:131–135
Parsons JA (1976) Parathyroid physiology and the skeleton. In: Bourne GH (ed) The biochemistry and physiology of bone, vol 4. Academic Press, New York, p 159
Herrman-Erlee MPM, Heersche JNM, Hekkelman JW, Gaillard PJ, Tregear GW, Parsons JA, Potts JT, Jr (1976) Effects on bonein vitro of bovine parathyroid hormone and synthetic fragments representing residues 1–34, 2–34 and 3–34. Endocrine Res Comm 3:21–35
Weisbrode SE, Capen CC, Nagode LA (1974) Effects of parathyroid hormone on bone of thyroparathyroidectomized rats Am J Pathol 75:529–536
Raisz LG, Kream BE (1981) Hormonal control of skeletal growth. Ann Rev Physiol 43:225–238
Robison R (1923) The possible significance of hexosphosphate esters in ossification. Biochem J 17:283–293
Fell HB, Robison R (1929) The growth, development and phosphatase activity of embryonic avian femora and limb buds cultivatedin vitro. Biochem J 23:767–787
Bourne GH (1972) Phosphatase and calcification. In: Bourne GH (ed) The biochemistry and physiology of bone, vol 2. Academic Press, New York, p 79
Osdoby P, Caplan AI (1981) Characterization of a bone-specific alkaline phosphatase in chick limb mesenchymal cell cultures. Develop Biol 86:136–146
Luben RA, Wong GL, Cohn DV (1976) Biochemical characterization with parathormone and calcitonin of isolated bone cells: Provisional identification of osteoclasts and osteoblasts. Endocrinology 99:526–534
Majeska RJ, Rodan SB, Rodan GA (1980) Parathyroid hormone-responsive clonal cell lines from rat osteosarcoma. Endocrinology 107:1494–1503
Osdoby P, Caplan AI (1981) First bone formation in the developing chick limb. Develop Biol 86:147–156
Peck WA, Birge AJ, Fedak SA (1964) Bone cells: Biochemical and biological studies after enzymatic isolation. Science 146:1476–1477
Brunk CF, Jones KC, James TW (1979) Assay for nanogram quantities of DNA in cellular homogenates. Anal Biochem 92:497–500
Lowry OH, Roberts NR, Wu M, Hixon WS, Crawford EJ (1954) The quantitative histochemistry of the brain. II. Enzyme measurements. J Biol Chem 207:19–37
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Kaplow LS (1963) Cytochemistry of leukocyte alkaline phosphatase. Am J Clin Pathol 39:439–449
McComb RB, Bowers GN, Posen S (1979) Alkaline phosphatase. Plenum Press, New York
Van Belle H (1972) Kinetics and inhibition of alkaline phosphatases from canine tissue. Biochim Biophys Acta 289:158–168
Majeska RJ, Rodan GA (1982) Alkaline phosphatase inhibition by parathyroid hormone and isoproterenol in a clonal rat osteosarcoma cell line. Possible mediation by cyclic AMP. Calcif Tissue Int 34:59–66
Woolfe CM (1968) Principles of biometry. D van Nostrand, Princeton
Wong GL, Luben RA, Cohn DV (1977) 1,25-dihydroxycho-lecalciferol and parathormone: Effects on isolated osteoclast-like and osteoblast-like cells. Science 197:663–665
Kumegawa M, Ikeda E, Tanaka S, Haneji T, Yora T, Sakagishi Y, Minami N, Hiramatsu M (1984) The effects of prostaglandin E2, parathyroid hormone, 1,25 dihydroxycholecalciferol, and cyclic nucleotide analogs on alkaline phosphatase activity in osteoblastic cells. Calcif Tissue Int 36:72–76
MacDonald BR, Gallagher JA, Ahnfelt Ronne I, Russell RGG (1984) PTH stimulates the production of prostaglandins by cells derived from human bone. Calcif Tissue Intl 36:465
Turner RT, Puzas JE, Forte MD, Lester GE, Gray TK, Howard GA, Baylink DJ (1980)In vitro synthesis of 1,25-dihydroxycholecalciferol and 24, 25-dihydroxycholecalciferol by isolated calvarial cells. Proc Natl Acad Sci USA 77:5720–5724
Howard GA, Turner RT, Sherrard DJ, Baylink DJ (1981) Human bone cells in culture metabolize 25-hydroxyvitamin D3 to 1,25-dihydroxyvitamin D3 and 24,25-dihydroxyvitamin D3. J Biol Chem 256:7738–7740
Mulkins MA, Mangolagas SC, Deftos LJ, Sussman HH (1983) 1,25-dihydroxyvitamin D3 increases bone alkaline phosphatase isoenzyme levels in human osteogenic sarcoma cells. J Biol Chem 258:6219–6225
Mangolagas SC, Burton DW, Deftos LJ (1981) 1,25-dihydroxyvitamin D3 stimulates the alkaline phosphatase activity of osteoblast-like cells. J Biol Chem 256:7115–7117
Majeska RJ, Rodan GA (1982) The effect of 1,25(OH)2D3 on alkaline phosphatase in osteoblastic osteosarcoma cells. J Biol Chem 257:3362–3365
Kurihara N, Ikeda K, Hakeda Y, Tsunoi M, Maeda N, Kumegawa M (1984) Effect of 1,25-dihydroxyvitamin D3 on alkaline phosphatase activity and collagen synthesis in osteoblastic cells, MC3T3-E1. Biochem Biophys Res Comm 119:767–771
Horwitz, KB, Koseki Y, McGuire WL (1978) Estrogen control of progesterone receptor in human breast cancer: Role of estradiol and antiestrogen. Endocrinology 103:1742–1751
Yee JA, Shew RL, Kenny AD, Pang PKT (1983) Effects of oxidized synthetic parathyroid hormone on bonein vitro. Calcif Tissue Int 35:689
Green H, Kehinde O (1975) An established preadipose cell line and its differentiation in culture. II. Factors affecting the adipose conversion. Cell 5:19–27
Linkhart TA, Clegg CH, Hauschka SD (1981) Myogenic differentiation in permanent clonal mouse myoblast cell lines: Regulation by macromolecular growth factors in culture medium. Develop Biol 86:19–30
Nakatani Y, Tsunoi M, Hakeda Y, Kurihara N, Fujita K, Kumegawa M (1984) Effects of parathyroid hormone on cAMP production and alkaline phosphatase activity in osteoblastic clone MC3T3-E1 cells. Biochem Biophys Res Comm 123:894–898
Felix R, Fleisch H (1979) Increase in alkaline phosphatase activity in calvaria cells cultured with diphosphonates. Biochem J 183:73–81
Chen TL, Cone CM, Feldman D (1983) Glucoccorticoid modulation of cell proliferation in cultured osteoblast-like cells: Differences between rat and mouse. Endocrinology 112:1739–1745
Chen TL, Feldman D (1984) Modulation of PTH-stimulated cyclic AMP in cultured rodent bone cells: The effects of 1,25(OH)2 vitamin D3 and its interaction with glucocorticoids. Calcif Tissue Intl 36:580–585
Peck WA, Klahr S (1979) Cyclic nucleotides in bone and mineral metabolism. In: Greengard P, Robison GA (eds) Advances in cyclic nucleotide research, vol 11. Raven Press, New York, p 89
Koyama H, Kato R, Ono T (1972) Induction of alkaline phosphatase by cyclic AMP or its dibutyryl derivative in a hybrid line between mouse and Chinese hamster in culture. Biochem Biophys Res Comm 46:305–311
Nose K, Katsuta H (1974) Induction of alkaline phosphatase activity by dibutyryl adenosine 3′, 5′-cyclic monophosphate in aneuploid rat liver cells. Exp Cell Res 87:8–14
Firestone GL, Heath EC (1981) The cyclic AMP-mediated induction of alkaline phosphatase in mouse L-cells. J Biol Chem 256:1396–1403
Burch WM, Lebovitz HE (1982)In vitro stimulation of alkaline phosphatase activity in immature embryonic chick pelvic cartilage by adenosine 3′,5′-monophosphate. J Cell Biol 93:338–342
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Yee, J.A. Stimulation of alkaline phosphatase activity in cultured neonatal mouse calvarial bone cells by parathyroid hormone. Calcif Tissue Int 37, 530–538 (1985). https://doi.org/10.1007/BF02557837
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF02557837