Calcified Tissue International

, Volume 82, Issue 2, pp 148–154 | Cite as

The Lipogenic Gene Spot 14 is Activated in Bone by Disuse yet Remains Unaffected by a Mechanical Signal Anabolic to the Skeleton

  • Jizu Zhi
  • Gang Xu
  • Clinton T. Rubin
  • Michael Hadjiargyrou


There is increasing evidence of the interaction of fat and bone metabolism and the role mechanical signals may have in regulating the adaptation of these tissues. The rat hindlimb suspension model of disuse osteoporosis was used to identify genes differentially expressed relative to normal weight-bearing bones and whether the relative expression of these genes is sensitive to anabolic mechanical stimuli. Ten days of hindlimb suspension suppressed percent labeled surface and bone volume/trabecular volume of the proximal tibia by 46% and 69%, respectively, compared to controls. Differential display polymerase chain reaction (DD-PCR) and Northern blot analysis identified and verified, respectively, that expression of Spot 14 (S14), an important gene in lipogenesis, was upregulated fourfold in tibiae of tail-suspended animals compared to long-term controls. Anabolic mechanical stimulation (45 Hz, 10 min/day at 0.25 g) did not show a statistically significant effect on S14 expression. These results indicate a potential role for lipogenic genes during bone loss caused by disuse, further supporting a link between bone and fat tissue, and, considering the insensitivity of these genes to mechanical signals which promote bone formation in the skeleton, the independence of resorptive and formative processes in bone.


Osteoporosis Disuse Mechanical stimulation Spot 14 


  1. 1.
    Lang T, LeBlanc A, Evans H, Lu Y, Genant H, Yu A (2004) Cortical, trabecular bone mineral loss from the spine and hip in long-duration spaceflight. J Bone Miner Res 19:1006–1012PubMedCrossRefGoogle Scholar
  2. 2.
    Mackay DL, Tesar PJ, Liang LN, Haynesworth SE (2006) Characterizing medullary and human mesenchymal stem cell-derived adipocytes. J Cell Physiol 207:722–728PubMedCrossRefGoogle Scholar
  3. 3.
    Lee NK, Sowa H, Hinoi E, Ferron M, Ahn JD, Confavreux C, Dacquin R, Mee PJ, McKee MD, Jung DY, Zhang Z, Kim JK, Mauvais-Jarvis F, Ducy P, Karsenty G (2007) Endocrine regulation of energy metabolism by the skeleton. Cell 130:456–469PubMedCrossRefGoogle Scholar
  4. 4.
    Rubin CT, Capilla E, Luu YK, Busa B, Crawford H, Nolan DJ, Mittal V, Rosen CJ, Pessin JE, Judex S (2007) Adipogenesis is inhibited by brief, daily exposure to high-frequency, extremely low-magnitude mechanical signals. Proc Natl Acad Sci USA 104:17879–17884PubMedCrossRefGoogle Scholar
  5. 5.
    Jump DB, Oppenheimer JH (1985) High basal expression and 3,5,3’-triiodothyronine regulation of messenger ribonucleic acid S14 in lipogenic tissues. Endocrinology 117:2259–2266PubMedCrossRefGoogle Scholar
  6. 6.
    Seelig S, Liaw C, Towle HC, Oppenheimer JH (1981) Thyroid hormone attenuates and augments hepatic gene expression at a pretranslational level. Proc Natl Acad Sci USA 78:4733–4737PubMedCrossRefGoogle Scholar
  7. 7.
    Freake HC, Oppenheimer JH (1987) Stimulation of S14 mRNA and lipogenesis in brown fat by hypothyroidism, cold exposure, and cafeteria feeding: evidence supporting a general role for S14 in lipogenesis and lipogenesis in the maintenance of thermogenesis. Proc Natl Acad Sci USA 84:3070–3074PubMedCrossRefGoogle Scholar
  8. 8.
    Kinlaw WB, Perez-Castillo AM, Fish LH, Mariash CN, Schwartz HL, Oppenheimer JH (1987) Interaction of dietary carbohydrate and glucagon in regulation of rat hepatic messenger ribonucleic acid S14 expression: role of circadian factors and 3’,5’-cyclic adenosine monophosphate. Mol Endocrinol 1:609–613PubMedCrossRefGoogle Scholar
  9. 9.
    Perez-Castillo A, Schwartz HL, Oppenheimer JH (1987) Rat hepatic mRNA-S14 and lipogenic enzymes during weaning: role of S14 in lipogenesis. Am J Physiol Endocrinol Metab 253:E536–E542Google Scholar
  10. 10.
    Sudo Y, Goto Y, Mariash CN (1993) Location of a glucose-dependent response region in the rat S14 promoter. Endocrinology 133:1221–1229PubMedCrossRefGoogle Scholar
  11. 11.
    Zhu Q, Anderson GW, Mucha GT, Parks EJ, Metkowski JK, Mariash CN (2005) The Spot 14 protein is required for de novo lipid synthesis in the lactating mammary gland. Endocrinology 146:3343–3350PubMedCrossRefGoogle Scholar
  12. 12.
    Zhu Q, Mariash A, Margosian MR, Gopinath S, Fareed MT, Anderson GW, Mariash CN (2001) Spot 14 gene deletion increases hepatic de novo lipogenesis. Endocrinology 142:4363–4370PubMedCrossRefGoogle Scholar
  13. 13.
    Sanchez-Rodriguez J, Kaninda-Tshilumbu JP, Santos A, Perez-Castillo A (2005) The spot 14 protein inhibits growth and induces differentiation and cell death of human MCF-7 breast cancer cells. Biochem J 390:57–65PubMedCrossRefGoogle Scholar
  14. 14.
    Kinlaw WB, Church JL, Harmon J, Mariash CN (1995) Direct evidence for a role of the “spot 14” protein in the regulation of lipid synthesis. J Biol Chem 270:16615–16618PubMedCrossRefGoogle Scholar
  15. 15.
    Kinlaw WB, Tron P, Friedmann AS (1992) Nuclear localization and hepatic zonation of rat “spot 14” protein: immunohistochemical investigation employing anti-fusion protein antibodies. Endocrinology 131:3120–3122PubMedCrossRefGoogle Scholar
  16. 16.
    LaFave LT, Augustin LB, Mariash CN (2006) S14: insights from knockout mice. Endocrinology 147:4044–4047PubMedCrossRefGoogle Scholar
  17. 17.
    Felson DT, Zhang Y, Hannan MT, Anderson JJ (1993) Effects of weight and body mass index on bone mineral density in men and women: the Framingham study. J Bone Miner Res 8:567–573PubMedGoogle Scholar
  18. 18.
    Ravn P, Cizza G, Bjarnason NH, Thompson D, Daley M, Wasnich RD, McClung M, Hosking D, Yates AJ, Christiansen C (1999) Low body mass index is an important risk factor for low bone mass and increased bone loss in early postmenopausal women. Early Postmenopausal Intervention Cohort (EPIC) study group. J Bone Miner Res 14:1622–1627PubMedCrossRefGoogle Scholar
  19. 19.
    Tremollieres FA, Pouilles JM, Ribot C (1993) Vertebral postmenopausal bone loss is reduced in overweight women: a longitudinal study in 155 early postmenopausal women. J Clin Endocrinol Metab 77:683–686PubMedCrossRefGoogle Scholar
  20. 20.
    Morey-Holton ER, Globus RK (2002) Hindlimb unloading rodent model: technical aspects. J Appl Physiol 92:1367–1377PubMedCrossRefGoogle Scholar
  21. 21.
    Rubin C, Xu G, Judex S (2001) The anabolic activity of bone tissue, suppressed by disuse, is normalized by brief exposure to extremely low-magnitude mechanical stimuli. FASEB J 15:2225–2229PubMedCrossRefGoogle Scholar
  22. 22.
    Erben RG (1997) Embedding of bone samples in methylmethacrylate: an improved method suitable for bone histomorphometry, histochemistry, and immunohistochemistry. J Histochem Cytochem 45:307–313PubMedGoogle Scholar
  23. 23.
    Parfitt AM, Drezner MK, Glorieux FH, Kanis JA, Malluche H, Meunier PJ, Ott SM, Recker RR (1987) Bone histomorphometry: standardization of nomenclature, symbols, and units. Report of the ASBMR Histomorphometry Nomenclature Committee. J Bone Miner Res 2:595–610PubMedCrossRefGoogle Scholar
  24. 24.
    Hadjiargyrou M, Halsey MF, Ahrens W, Rightmire EP, McLeod KJ, Rubin CT (1998) Cloning of a novel cDNA expressed during the early stages of fracture healing. Biochem Biophys Res Commun 249:879–884PubMedCrossRefGoogle Scholar
  25. 25.
    Liang P, Pardee AB (1992) Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257:967–971PubMedCrossRefGoogle Scholar
  26. 26.
    Hadjiargyrou M, Ahrens W, Rubin CT (2000) Temporal expression of the chondrogenic and angiogenic growth factor CYR61 during fracture repair. J Bone Miner Res 15:1014–1023PubMedCrossRefGoogle Scholar
  27. 27.
    Zhi J, Sommerfeldt DW, Rubin CT, Hadjiargyrou M (2001) Differential expression of neuroleukin in osseous tissues and its involvement in mineralization during osteoblast differentiation. J Bone Miner Res 16:1994–2004PubMedCrossRefGoogle Scholar
  28. 28.
    Judex S, Zhong N, Squire ME, Ye K, Donahue LR, Hadjiargyrou M, Rubin CT (2005) Mechanical modulation of molecular signals which regulate anabolic and catabolic activity in bone tissue. J Cell Biochem 94:982–994PubMedCrossRefGoogle Scholar
  29. 29.
    Zayzafoon M, Gathings WE, McDonald JM (2004) Modeled microgravity inhibits osteogenic differentiation of human mesenchymal stem cells and increases adipogenesis. Endocrinology 145:2421–2432PubMedCrossRefGoogle Scholar
  30. 30.
    Burkhardt R, Kettner G, Bohm W, Schmidmeier M, Schlag R, Frisch B, Mallmann B, Eisenmenger W, Gilg T (1987) Changes in trabecular bone, hematopoiesis and bone marrow vessels in aplastic anemia, primary osteoporosis, and old age: a comparative histomorphometric study. Bone 8:157–164PubMedCrossRefGoogle Scholar
  31. 31.
    Meunier P, Aaron J, Edouard C, Vignon G (1971) Osteoporosis and the replacement of cell populations of the marrow by adipose tissue. A quantitative study of 84 iliac bone biopsies. Clin Orthop Relat Res 80:147–154PubMedCrossRefGoogle Scholar
  32. 32.
    Wronski TJ, Walsh CC, Ignaszewski LA (1986) Histologic evidence for osteopenia and increased bone turnover in ovariectomized rats. Bone 7:119–123PubMedCrossRefGoogle Scholar
  33. 33.
    Minaire P, Neunier P, Edouard C, Bernard J, Courpron P, Bourret J (1974) Quantitative histological data on disuse osteoporosis: comparison with biological data. Calcif Tissue Res 17:57–73PubMedCrossRefGoogle Scholar
  34. 34.
    Wang GJ, Sweet DE, Reger SI, Thompson RC (1977) Fat-cell changes as a mechanism of avascular necrosis of the femoral head in cortisone-treated rabbits. J Bone Joint Surg Am 59:729–735PubMedGoogle Scholar
  35. 35.
    Chou WY, Cheng YS, Ho CL, Liu ST, Liu PY, Kuo CC, Chang HP, Chen YH, Chang GG, Huang SM (2007) Human spot 14 protein interacts physically and functionally with the thyroid receptor. Biochem Biophys Res Commun 357:133–138PubMedCrossRefGoogle Scholar
  36. 36.
    Bassett JH, O’Shea PJ, Sriskantharajah S, Rabier B, Boyde A, Howell PG, Weiss RE, Roux JP, Malaval L, Clement-Lacroix P, Samarut J, Chassande O, Williams GR (2007) Thyroid hormone excess rather than thyrotropin deficiency induces osteoporosis in hyperthyroidism. Mol Endocrinol 21:1095–1107PubMedCrossRefGoogle Scholar
  37. 37.
    Murphy E, Williams GR (2004) The thyroid and the skeleton. Clin Endocrinol (Oxf) 61:285–298CrossRefGoogle Scholar
  38. 38.
    Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147PubMedCrossRefGoogle Scholar
  39. 39.
    Beresford JN, Bennett JH, Devlin C, Leboy PS, Owen ME (1992) Evidence for an inverse relationship between the differentiation of adipocytic and osteogenic cells in rat marrow stromal cell cultures. J Cell Sci 102(Pt 2):341–351PubMedGoogle Scholar
  40. 40.
    Nuttall ME, Patton AJ, Olivera DL, Nadeau DP, Gowen M (1998) Human trabecular bone cells are able to express both osteoblastic and adipocytic phenotype: implications for osteopenic disorders. J Bone Miner Res 13:371–382PubMedCrossRefGoogle Scholar
  41. 41.
    Dorheim MA, Sullivan M, Dandapani V, Wu X, Hudson J, Segarini PR, Rosen DM, Aulthouse AL, Gimble JM (1993) Osteoblastic gene expression during adipogenesis in hematopoietic supporting murine bone marrow stromal cells. J Cell Physiol 154:317–328PubMedCrossRefGoogle Scholar
  42. 42.
    Meyer MH, Etienne W, Meyer RA Jr (2004) Altered mRNA expression of genes related to nerve cell activity in the fracture callus of older rats: a randomized, controlled, microarray study. BMC Musculoskelet Disord 5:24PubMedCrossRefGoogle Scholar
  43. 43.
    Chawla A, Boisvert WA, Lee CH, Laffitte BA, Barak Y, Joseph SB, Liao D, Nagy L, Edwards PA, Curtiss LK, Evans RM, Tontonoz P (2001) A PPAR gamma-LXR-ABCA1 pathway in macrophages is involved in cholesterol efflux and atherogenesis. Mol Cell 7:161–171PubMedCrossRefGoogle Scholar
  44. 44.
    Laffitte BA, Joseph SB, Walczak R, Pei L, Wilpitz DC, Collins JL, Tontonoz P (2001) Autoregulation of the human liver X receptor alpha promoter. Mol Cell Biol 21:7558–7568PubMedCrossRefGoogle Scholar
  45. 45.
    Repa JJ, Liang G, Ou J, Bashmakov Y, Lobaccaro JM, Shimomura I, Shan B, Brown MS, Goldstein JL, Mangelsdorf DJ (2000) Regulation of mouse sterol regulatory element-binding protein-1c gene (SREBP-1c) by oxysterol receptors, LXRalpha and LXRbeta. Genes Dev 14:2819–2830PubMedCrossRefGoogle Scholar
  46. 46.
    Foretz M, Pacot C, Dugail I, Lemarchand P, Guichard C, Le Liepvre X, Berthelier-Lubrano C, Spiegelman B, Kim JB, Ferre P, Foufelle F (1999) ADD1/SREBP-1c is required in the activation of hepatic lipogenic gene expression by glucose. Mol Cell Biol 19:3760–3768PubMedGoogle Scholar
  47. 47.
    Hummasti S, Laffitte BA, Watson MA, Galardi C, Chao LC, Ramamurthy L, Moore JT, Tontonoz P (2004) Liver X receptors are regulators of adipocyte gene expression but not differentiation: identification of apoD as a direct target. J Lipid Res 45:616–625PubMedCrossRefGoogle Scholar
  48. 48.
    Teran-Garcia M, Adamson AW, Yu G, Rufo C, Suchankova G, Dreesen TD, Tekle M, Clarke SD, Gettys TW (2007) Polyunsaturated fatty acid suppression of fatty acid synthase (FASN): evidence for dietary modulation of NF-Y binding to the Fasn promoter by SREBP-1c. Biochem J 402:591–600PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Jizu Zhi
    • 1
  • Gang Xu
    • 1
  • Clinton T. Rubin
    • 1
  • Michael Hadjiargyrou
    • 1
  1. 1.Department of Biomedical EngineeringStony Brook UniversityStony BrookUSA

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