Transgenic Research

, Volume 3, Issue 2, pp 127–133 | Cite as

Regulation of insulin-like growth factor-l and binding protein-3 expression in oMtla-oGH transgenic mice

  • Jesse C. Chow
  • James D. Murray
  • Daniel Pomp
  • Ransom L. Baldwin
  • Christopher C. Calvert
  • Anita M. Oberbauer


Growth hormone (GH)-transgenic mice provide a model for studying hormonal regulation of gene products responsible for efficient lean growth. Insulin-like growth factor-I (IGF-I) and IGF binding protein-3 (BP-3) are two products involved in mediating the growth promoting actions of GH. Mice carrying the ovine metallothionein la-ovine growth hormone (oMtla-oGH) transgene were used to study GH regulation of IGF-I and PB-3 expression because these mice do not exhibit elevated basal oGH levels without transgene stimulation by exogenous zinc. C57B1/6XCBA mice with (transgenic=TG) and without (control=C) the oMtla-oGH transgene were activated (+Zn) or inactivated (-Zn) by the addition or removal of 25 mM zinc sulfate in the drinking water. Plasma IGF-I and BP-3 levels were determined by radioimmunoassay and western ligand blotting, respectively. Hepatic IGF-I and BP-3 mRNA levels were determined by slot-blot analysis. TG+Zn mice had higher plasma IGF-I (p<0.05) and hepatic IGF-I mRNA (p<0.05) levels as compared to TG-Zn, C+Zn and C-Zn mice. Plasma IGF-I and hepatic IGF-I mRNA levels in TG-Zn mice were not different from C+Zn and C-Zn mice. Removal of Zn decreased hepatic IGF-I mRNA levels to C levels in TG mice. Plasma BP-3 and hepatic BP-3 mRNA levels in TG+Zn mice were increased (p<0.05) as compared to TG-Zn, C-Zn and C+Zn. Plasma BP-3 and hepatic BP-3 mRNA levels did not differ between TG-Zn, C-Zn and C+Zn mice. Expression of the transgene also increased the level of plasma BP-3 during pregnancy as compared to that observed for pregnant C mice. We conclude that oGH regulates IGF-I and BP-3 expression in the oMtla-oGH transgenic mouse model system.


IGF-I BP-3 GH transgenic mouse pregnancy 


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  1. Adamo, M.L., Shao, Z., Lanau, F., Chen, J.C., Clemmons, D.R., Roberts, C.T., Jr., LeRoith, D. and Fontana, J.A. (1992). Insulin-like growth factor-I and retinoic acid modulation of IGF-binding proteins (IGFBPs): IGFBP-2,-3, and-4 gene expression and protein secretion in a breast cancer cell line.Endocrinol. 131, 1858–66.Google Scholar
  2. Albiston, A.L. and Herrington, A.C. (1992) Tissue distribution and regulation of insulin-like growth factor (IGF)-binding protein-3 messenger ribonucleic acid (mRNA) in the rat: comparison with IGF-I mRNA expression.Endocrinol. 130, 492–502.Google Scholar
  3. Bar, R.S., Boes, M., Dake, B.L., Sandra, A., Bayne, M., Cascieri, M., and Booth, B.A. (1990) Tissue localization of perfused endothelial cell IGF binding protein is markedly altered by association with IGF-I.Endocrinol. 127, 3243–45.Google Scholar
  4. Baxter, R.C., and Martin, J.L. (1989) Binding proteins for insulin-like growth factors: structure, regulation and function.Prog. Growth Factor Res. 1, 49–68.Google Scholar
  5. Breier, B.H., Gallaher, B.W. and Gluckman, P.D. (1991) Radioimmunoassay for insulin-like growth factor-I: solutions to some potential problems and pitfalls.J. Endo. 128, 347–57.Google Scholar
  6. Chomczynski, P. and Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenolchloroform extraction.Anal. Biochem. 162, 156–9.Google Scholar
  7. Clemmons, D.R., Thissen, J.P., Maes, M., Ketelslegers, J.M. and Underwood, L.E. (1989) Insulin-like growth factor infusion into hypophysectomized or protein-deprived rats induces specific IGF binding proteins in serum.Endocrinol. 125, 2967–72.Google Scholar
  8. Cohick, W.S. and Clemmons, D.R. (1991) Regulation of insulinlike growth factor binding protein synthesis and secretion in a bovine epithelial cell line.Endocrinol. 129, 1347–54.Google Scholar
  9. Copeland, K.C., Underwood, L.E. and Van Wyk, J.J. (1980) Induction of immunoreactive somatomedin-C in human serum by growth hormone: dose response relationships and effect on chromatographic profiles.J. Clin. Endo. & Metab. 50, 690–7.Google Scholar
  10. Davenport, M.L., Clemmons, D.R., Miles, M.V., Camacho-Hubner, C., D'Ercole, J.A. and Underwood, L.E. (1990) Regulation of serum insulin-like growth factor-I (IGF-I) and IGF binding proteins during rat pregnancy.Endocrinol. 127 1278–85.Google Scholar
  11. Davenport, M.L., Pucilowska, J., Clemmons, D.R., Lundblad, R., Spencer, J.A. and Underwood, L.E. (1992) Tissue-specific expression of insulin-like growth factor binding protein-3 protease activity during rat pregnancy.Endocrinol. 130, 2505–12.Google Scholar
  12. De Mellow, J.S.M. and Baxter, R.C. (1988) Growth hormone dependent insulin-like growth factor binding protein both inhibits and potentiates IGF-I stimulated DNA synthesis in skin fibroblasts.Biochem. Biophys. Res. Commun. 156, 199–204.Google Scholar
  13. Gargosky, S.E., Nanto-Salonen, K., Tapanainen, P. and Rosenfield, R.G. (1993) Pregnancy in growth hormone-deficient rats: assessment of insulin-like growth factors (IGFs), IGF-binding proteins (IGFBPs) and IGFBP protease activity,J. Endo. 136, 479–89.Google Scholar
  14. Guler, H.P., Eckardt, K.U., Zapf, J., Bauer, C. and Froesch, E.R. (1989) Insulin-like growth factor I increases glomerular filtration rate and renal plasma flow in man.Acta Endocrinol. 121, 101–6.Google Scholar
  15. Gunning, P., Ponte, P., Okayama, H., Engel, J., Blau, H. and Kedes, L. (1983) Isolation and characterization of full length cDNA clones for human α-, β-, and γ-actin mRNA's: skeletal but not cytoplasmic have an amino terminal cysteine that is subsequently removed.Mol. Cell. Biol. 3, 787–95.Google Scholar
  16. Hammreman, M.R. (1989) The growth hormone-insulin-like growth factor axis in kidney.Am. J. Physiol. 257, F503–14.Google Scholar
  17. Hodgkinson, S.C., Moore, L., Napier, J.R., Davis, S.R., Bass, J.J., and Gluckman, P.D. (1989) Characterization of IGFBPs in ovine tissue fluids.J. Endo. 120, 429–38.Google Scholar
  18. Hossenlopp, P., Seurin, D., Segovia-Quinson, B., Hardouin, S. and Binoux, M. (1986) Analysis of serum insulin-like factor binding proteins using western blotting: use of the method for titration of the binding proteins and competitive binding studies.Anal. Biochem. 154, 138–43.Google Scholar
  19. Laemmli, U.K. (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4.Nature 227, 680–5.Google Scholar
  20. Lowe, W.L., Jr. (1991) Biological actions of insulin-like growth factors. In LeRoith, D. ed.,Insulin-like Growth Factors: Molecular and Cellular Aspects, pp. 49–85. Boca Raton: CRC Press.Google Scholar
  21. Mathews, L.S., Norstedt, G. and Palmiter, R.D. (1986) Regulation of insulin-like growth factor I gene expression by growth hormone.Proc. Natl Acad. Sci. USA 83, 9343–7.Google Scholar
  22. McCarthy, T.L., Centrella, M. and Canalis, E. (1990) Cyclic AMP induces insulin-like growth factor I synthesis in osteoblast-enriched cultures.J. Biol. Chem. 265, 15353–6.Google Scholar
  23. Nilsson, A., Carlsson, B., Isgaard, J., Isaksson, O.G. and Rymo, L. (1990) Regulation by GH of insulin-like growth factor-I mRNA expression in rat epiphyseal growth plate as studied within-situ hybridization.J. Endocrinol. 129, 67–74.Google Scholar
  24. Oberbauer, A.M., Currier, T.A., Nancarrow, C.D., Ward, K.A., and Murray, J.D. (1992) Linear bone growth of oMtla-oGH transgenic male mice.Am. J. Physiol. 262, E936–42.Google Scholar
  25. Palmiter, R.D., Norstedt, G., Gelinas, R.E., Hammer, R.E. and Brinster, R.L. (1983) Metallothionein-human GH fusion genes stimulate growth of mice.Science 222, 809–14.Google Scholar
  26. Pomp, D., Murray, J.D. and Medrano, J.F. (1991) Single day detection of transgenic mice by PCR of toe-clips.Mouse Genome 89, 279.Google Scholar
  27. Pomp, D., Nancarrow, C.D., Ward, K.A. and Murray, J.D. (1992) Growth, feed efficiency and body composition of transgenic mice expressing a sheep metallothionein 1a-sheep growth hormone fusion gene.Livest. Prod. Sci. 31, 335–50.Google Scholar
  28. Prewitt, T.E., D'Ercole, J.A., Switzer, B.R. and Van Wyk, J.J. (1982) Relationship of serum immunoreactive somatomedian-C to dietary protein and energy in growth rats.J. Nutr. 112, 144–50.Google Scholar
  29. Salmon, W.D. and Daughaday, W.H. (1957) A hormonally controlled serum factor which stimulates sulfate incorporation by cartilagein vitro.J. Lab. Clin. Med. 49, 825–36.Google Scholar
  30. Sambrook, J., Fritsch, E.F. and Maniatis, T. ed. (1989)Molecular Cloning: a Laboratory Manual,1, 7.37–7.79. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press.Google Scholar
  31. Searle, S.R. (1987)Linear Models for Unbalanced Data New York: John Wiley & Sons.Google Scholar
  32. Shanahan, C.M., Rigby, N.W., Murray, J.D., Marshall, J.T., Townrow, C.A., Nancarrow, C.D. and Ward, K.A. (1989) Regulation of expression of a sheep metallothionein 1a-sheep growth hormone fusion gene in transgenic mice.Mol. Cell. Biol. 9, 5473–9.Google Scholar
  33. Straus, D.S. and Takemoto, C.D. (1990) Effect of dietary protein deprivation on insulin-like growth factor (IGF)-I and-II, IGF binding protein-2, and serum albumin gene expression in rat.Endocrinol. 127, 1849–60.Google Scholar
  34. Yoon, J., Berry, S.A., Seelig, S. and Towle, H.C. (1990) An inducible nuclear factor binds to a growth hormone-regulated gene.J. Biol. Chem. 265, 19947–54.Google Scholar
  35. Zapf, J., Hauri, C., Waldvogel, M., Futo, E., Hasler, H., Binz, K., Guler, H.P., Schimid, C. and Froesch, E.R. (1989) Recombinant human insulin-like growth factor I induces its own specific carrier protein in hypophysectomized and diabetic rats.Proc. Natl Acad. Sci. USA 86, 3813–7.Google Scholar

Copyright information

© Chapman & Hall 1994

Authors and Affiliations

  • Jesse C. Chow
    • 1
  • James D. Murray
    • 1
    • 2
  • Daniel Pomp
    • 1
    • 3
  • Ransom L. Baldwin
    • 1
  • Christopher C. Calvert
    • 1
  • Anita M. Oberbauer
    • 1
  1. 1.Departments of Animal Science, School of Veterinary MedicineUniversity of California-DavisDavisUSA
  2. 2.Department of Reproduction, School of Veterinary MedicineUniversity of California-DavisDavisUSA
  3. 3.Oklahoma State UniversityStillwaterUSA

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