Anabolic or Catabolic Responses of MC3T3-E1 Osteoblastic Cells to Parathyroid Hormone Depend on Time and Duration of Treatment

Oxford University Press (OUP) - Tập 14 Số 9 - Trang 1504-1512 - 1999
Paul C. Schiller1, Gianluca D’Ippolito, Bernard A. Roos, G. A. Howard
1Geriatric Research, Education, and Clinical Center, and Research Service, Veterans Affairs Medical Center, and Department of Medicine, Miami, Florida 33125, USA.

Tóm tắt

Abstract We have investigated signaling (cAMP) and anabolic responses (mineralization of extracellular matrix [ECM]) to parathyroid hormone (PTH) in long-term (30 days) cultures of MC3T3-E1 cells, a murine model of osteoblast differentiation. Expression of PTH/PTH–related peptide receptor (PTH1R) mRNA is detected early and remains relatively constant for 2 weeks with somewhat higher levels observed during the second half of the culture period. In contrast to the relatively stable PTH1R mRNA expression, the cAMP response to PTH varies markedly with no response at day 5 and a marked response (80-fold versus control) by day 10. Responsiveness to PTH remains elevated with fluctuations of 30- to 80-fold stimulation throughout the remainder of the culture period. The timing and duration of PTH treatment to achieve in vitro mineralization of ECM was evaluated. When continuous PTH treatment was initiated before day 20, mineralization decreased. If continuous PTH treatment began on or after day 20, mineralization was unaffected. However, if treatment began on day 20 and then stopped on day 25, mineralization on day 30 was increased 5-fold. This mineralization response to intermittent PTH was confirmed in primary cultures of murine and human osteoblastic cells. These data provide a potential basis for understanding the differential responses to PTH (anabolic versus catabolic) and indicate the developmental temporal variance of anabolic and catabolic responses. Since cAMP signaling was relatively unchanged during this interval (day 10–30) and stimulation of adenylate cyclase only partially mimicked the PTH effect on increased mineralization, other signaling pathways are likely to be involved in order to determine the specific anabolic response to short-term PTH treatment during the differentiation process.

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Tài liệu tham khảo

Bauer, 1929, Studies of calcium phosphorus metabolism: Study of bone trabeculae as ready available reserve supply of calcium, J Exp Med, 49, 145, 10.1084/jem.49.1.145

Selye, 1932, On the stimulation of new-bone-formation with parathyroid extract, irradiated ergosterol, Endocrinology, 16, 547, 10.1210/endo-16-5-547

Reeve, 1976, Anabolic effect of low doses of a fragment of human parathyroid hormone on the skeleton in postmenopausal osteoporosis, Lancet, 1, 1035, 10.1016/S0140-6736(76)92216-9

Hock, 1988, Human parathyroid hormone (1–34) increases bone mass in ovariectomized and orchidectomized rats, Endocrinology, 122, 2899, 10.1210/endo-122-6-2899

Mosekilde, 1991, The anabolic effects of human parathyroid hormone (hPTH) on rat vertebral body mass are also reflected in the quality of bone, assessed by biomechanical testing: A comparison study between hPTH-(1–34) and hPTH-(1–84), Endocrinology, 129, 421, 10.1210/endo-129-1-421

Ibbotson, 1992, Contrasting effects of parathyroid hormone and insulin-like growth factor I in an aged ovariectomized rat model of postmenopausal osteoporosis, J Bone Miner Res, 7, 425, 10.1002/jbmr.5650070410

Hock, 1992, Effects of continuous and intermittent administration and inhibition of resorption on the anabolic response of bone to parathyroid hormone, J Bone Miner Res, 7, 65, 10.1002/jbmr.5650070110

Dempster, 1993, Anabolic actions of parathyroid hormone in bone, Endocr Rev, 14, 690

Wronski, 1993, Parathyroid hormone is more effective than estrogen or bisphosphonates for restoration of lost bone mass in ovariectomized rats, Endocrinology, 132, 823, 10.1210/endo.132.2.8425497

Reeve, 1993, Human parathyroid peptide treatment of vertebral osteoporosis, Osteoporos Int, 3, 199, 10.1007/BF01621906

Qi, 1995, A comparison of the anabolic effects of parathyroid hormone at skeletal sites with moderate and severe osteopenia in aged ovariectomized rats, J Bone Miner Res, 10, 948, 10.1002/jbmr.5650100616

Li, 1995, Parathyroid hormone monotherapy and cotherapy with antiresorptive agents restore vertebral bone mass and strength in aged ovariectomized rats, Bone, 16, 629, 10.1016/8756-3282(95)00115-T

MacDonald, 1986, Parathyroid hormone stimulates the proliferation of cells derived from human bone, Endocrinology, 118, 2445, 10.1210/endo-118-6-2445

Canalis, 1989, Insulin-like growth factor I mediates selective anabolic effects of parathyroid hormone in bone cultures, J Clin Invest, 83, 60, 10.1172/JCI113885

McCarthy, 1989, Parathyroid hormone enhances the transcript and polypeptide levels of insulin-like growth factor I in osteoblast-enriched cultures from fetal rat bone, Endocrinology, 124, 1247, 10.1210/endo-124-3-1247

Nishida, 1994, Increased bone formation by intermittent parathyroid hormone administration is due to the stimulation of proliferation and differentiation of osteoprogenitor cells in bone marrow, Bone, 15, 717, 10.1016/8756-3282(94)90322-0

Schmidt, 1995, Intermittent parathyroid hormone treatment increases osteoblast number, steady state messenger ribonucleic acid levels for osteocalcin, and bone formation in tibial metaphysis of hypophysectomized female rats, Endocrinology, 136, 5127, 10.1210/endo.136.11.7588250

Dobnig, 1995, Evidence that intermittent treatment with parathyroid hormone increases bone formation in adult rats by activation of bone lining cells, Endocrinology, 136, 3632, 10.1210/endo.136.8.7628403

Rouleau, 1990, Characterization of the major parathyroid hormone target cell in the endosteal metaphysis of rat long bones, J Bone Miner Res, 5, 1043, 10.1002/jbmr.5650051008

Jüppner, 1991, A G protein-linked receptor for parathyroid hormone and parathyroid hormone-related peptide, Science, 254, 1024, 10.1126/science.1658941

Abou-Samra, 1992, Expression cloning of a common receptor for parathyroid hormone and parathyroid hormone-related peptide from rat osteoblast-like cells: A single receptor stimulates intracellular accumulation of both cAMP and inositol trisphosphates and increases intracellular free calcium, Proc Natl Acad Sci USA, 89, 2732, 10.1073/pnas.89.7.2732

Lee, 1994, Parathyroid hormone induces sequential c-fos expression in bone cells in vivo: In situ localization of its receptor and c-fos messenger ribonucleic acids, Endocrinology, 134, 441, 10.1210/endo.134.1.8275957

Jobert, 1996, Characterization of alternatively spliced isoforms of the PTH/PTHrP receptor messenger RNA expressed in human kidney and bone cells, J Bone Miner Res, 11, S303

Joun, 1997, Tissue-specific transcription start sites and alternative splicing of the parathyroid hormone (PTH)/PTH-related peptide (PTHrP) receptor gene: A new PTH/PTHrP receptor splice variant that lacks the signal peptide, Endocrinology, 138, 1742, 10.1210/endo.138.4.5085

Isogai, 1996, Parathyroid hormone regulates osteoblast differentiation positively or negatively depending on the differentiation stages, J Bone Miner Res, 11, 1384, 10.1002/jbmr.5650111003

Hollnagel, 1997, Parathyroid hormone enhances early and suppresses late stages of osteogenic and chondrogenic development in a BMP-dependent mesenchymal differentiation system (C3H10T1/2), J Bone Miner Res, 12, 1993, 10.1359/jbmr.1997.12.12.1993

McCauley, 1996, PTH/PTHrP receptor is temporally regulated during osteoblast differentiation and is associated with collagen synthesis, J Cell Biochem, 61, 638, 10.1002/(SICI)1097-4644(19960616)61:4<638::AID-JCB18>3.0.CO;2-B

Kondo, 1997, Temporal changes of mRNA expression of matrix proteins and parathyroid hormone and parathyroid hormone-related protein (PTH/PTHrP) receptor in bone development, J Bone Miner Res, 12, 2089, 10.1359/jbmr.1997.12.12.2089

Bos, 1996, Expression of the parathyroid hormone receptor and correlation with other osteoblastic parameters in fetal rat osteoblasts, Calcif Tissue Int, 58, 95, 10.1007/BF02529730

Ishizuya, 1997, Parathyroid hormone exerts disparate effects on osteoblast differentiation depending on exposure time in rat osteoblastic cells, J Clin Invest, 99, 2961, 10.1172/JCI119491

Kodama, 1981, Establishment of a clonal osteogenic cell line from newborn mouse calvaria, Jpn J Oral Biol, 23, 899, 10.2330/joralbiosci1965.23.899

Quarles, 1992, Distinct proliferative and differentiated stages of murine MC3T3–E1 cells in culture: An in vitro model of osteoblast development, J Bone Miner Res, 7, 683, 10.1002/jbmr.5650070613

Schiller, 1992, Hormonal regulation of intercellular communication: Parathyroid hormone increases connexin 43 gene expression and gap-junctional communication in osteoblastic cells, Mol Endocrinol, 6, 1433

Howard, 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, 10.1016/S0021-9258(18)43337-6

Chomezynsky, 1987, Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction, Anal Biochem, 162, 156

Labarca, 1980, A simple, rapid, and sensitive DNA assay procedure, Anal Biochem, 102, 344, 10.1016/0003-2697(80)90165-7

Manolagas, 1981, 1,25-dihydroxyvitamin D3 stimulates the alkaline phosphatase activity of osteoblast-like cells, J Biol Chem, 256, 7115, 10.1016/S0021-9258(19)68933-7

Stanford, 1995, Rapidly forming apatitic mineral in an osteoblastic cell line (UMR 106–01 BSP), J Biol Chem, 270, 9420, 10.1074/jbc.270.16.9420

Halverson, 1979, Identification of hydroxyapatite crystals in synovial fluid, Arthritis Rheum, 22, 389, 10.1002/art.1780220412

Partridge, 1994, Signal transduction pathways mediating parathyroid hormone regulation of osteoblastic gene expression, J Cell Biochem, 55, 321, 10.1002/jcb.240550308

Leis, 1997, Phenotypic heterogeneity of osteoblast-like MC3T3–E1 cells: Changes of bradykinin-induced prostaglandin E2 production during osteoblast maturation, J Bone Miner Res, 12, 541, 10.1359/jbmr.1997.12.4.541

Schirmacher, 1992, Characterization of gap junctions between osteoblast-like cells in culture, Calcif Tissue Int, 51, 285, 10.1007/BF00334489

Civitelli, 1993, Connexin 43 mediates direct intercellular communication in human osteoblastic cell networks, J Clin Invest, 91, 1888, 10.1172/JCI116406

Donahue, 1995, Cell-to-cell communication in osteoblastic networks: Cell line-dependent hormonal regulation of gap junction function, J Bone Miner Res, 10, 881, 10.1002/jbmr.5650100609

Yamaguchi, 1994, Isolation and characterization of gap junctions in the osteoblastic MC3T3–E1 cell line, J Bone Miner Res, 9, 791, 10.1002/jbmr.5650090605

Jones, 1993, The incidence and size of gap junctions between the bone cells in rat calvaria, Anat Embryol, 187, 343, 10.1007/BF00185892

Steinberg, 1994, Connexin43 and connexin45 form gap junctions with different molecular permeabilities in osteoblastic cells, EMBO J, 13, 744, 10.1002/j.1460-2075.1994.tb06316.x

Minkoff, 1994, Gap junction proteins exhibit early and specific expression during intramembranous bone formation in the developing chick mandible, Anat Embryol, 190, 231, 10.1007/BF00234301

Schiller, 1997, PTH up-regulation of connexin 43 gene expression in osteoblasts depends on cell phenotype, J Bone Miner Res, 12, 2005, 10.1359/jbmr.1997.12.12.2005

Vander Molen, 1996, Gap junctional communication contributes to hormonal responsiveness in osteoblastic networks, J Biol Chem, 271, 12165, 10.1074/jbc.271.21.12165

Majeska, 1985, Glucocorticoid regulation of alkaline phosphatase in the osteoblastic osteosarcoma cell line ROS 17/2.8, Endocrinology, 116, 170, 10.1210/endo-116-1-170

Yang, 1997, Selective stimulation of apoptosis in trabecular osteoblasts and osteocytes of young rats treated with once daily hPTH 1–34 to increase bone mass, J Bone Miner Res, 12, S316

Fladmark, 1997, Fas/APO-1 (CD95)-induced apoptosis of primary hepatocytes is inhibited by cAMP, Biochem Biophys Res Commun, 232, 20, 10.1006/bbrc.1997.6214

Sabatini, 1996, Effects of parathyroid hormone and agonists of the adenylyl cyclase and protein kinase C pathways on bone cell proliferation, Bone, 18, 59, 10.1016/8756-3282(95)00427-0

Hilliker, 1996, Truncation of the amino terminus of PTH alters its anabolic activity on bone in vivo, Bone, 19, 469, 10.1016/S8756-3282(96)00230-X

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Oxford University Press (OUP)

Tập 14 Số 9

1504-1512

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