Skip to main content
SpringerLink
Log in

Real-time detection of insulin-like growth factor-1 stimulation of the MAC-T bovine mammary epithelial cell line

  • Published:
Endocrine Aims and scope Submit manuscript

Abstract

Binding of growth factors by cell-surface receptors is an essential means by which cell regulate normaltissue growth and differentiation. Exposure to growth factors is often transient, and our goal was to determine whether short-term exposure to insulin-like growth factor-1 (IGF-1) would lead to activation, assayed as cell proliferation, of mammary epithelial cells. The MAC-T cell line is an immortalized bovine mammary epithelial cell line, chosen as our model mammary cell line because of its known sensitivity to IGF-1. Using the Cytosensor Microphysiometer System, a biosensor capable of measuring extracellular acidification, we were able to measure activation of the cellsowing to IGF-1 addition in real time and found that peak acidification occurred in only 14 min. We show that this rapid response to IGF-1 is dose dependent and specific for IGF-1. A significant increase in [3H]thymidine incorporation by cells after a similar short-term exposure to IGF-1 suggests that the measured increase in extracellular acidification following IGF-1 addition is physiologically relevant. This technology offers a new, novel, and rapid means for the study of IGF-1 activity, as well as the screening of IGF-1 inhibitors, in mammary epithelial cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Cohick, W. S. (1998). J. Dairy Sci. 81, 1769–1777.

    Article  PubMed  CAS  Google Scholar 

  2. Hovey, R. C., McFadden, T. B., and Akers, R. M. (1998). J. Mam. Gland Biol. Neoplasia 4, 53–68.

    Article  Google Scholar 

  3. Kleinberg, D. L. (1998). Breast Cancer Res Tr. 47, 201–208.

    Article  CAS  Google Scholar 

  4. Akers, R. M., McFadden, T. B., Purup, M., Vestergaard, M., Sejrsen, S., and Capuco, A. V. (2000). J. Mam. Gland Biol. Neoplasia 5, 43–51.

    Article  CAS  Google Scholar 

  5. Forsyth, I. A., Gabai, G., and Morgan, G. (1999). J. Dairy Res. 66, 35–44.

    Article  PubMed  CAS  Google Scholar 

  6. Weber, M. S., Purup, S., Vestergaard, M., Ellis, S. E., Sondergard-Andersen, J., Akers, R. M., and Sejrsen, K. (1999). J. Endocrinol. 161, 365–373.

    Article  PubMed  CAS  Google Scholar 

  7. Weber, M. S., Purup, S., Vestergaard, M., Akers, R. M., and Sejrsen, K. (2000). J. Dairy Sci. 83, 30–37.

    Article  PubMed  CAS  Google Scholar 

  8. Gill, G. N. (1990). In: Peptide growth factors and their receptors I Sporn, M. B. and Roberts, A. B. (eds.), Springer-Verlag: New York.

    Google Scholar 

  9. Blakesley, V. A., Scrimgeour, A., Esposito, D., and LeRoith, D. (1996). Cytokine Growth Factor Res. 7, 153–159.

    Article  CAS  Google Scholar 

  10. LeRoith, D., Werner, H., Beitner-Johnson, D., and Roberts, J. C. T. (1995). Endocr. Rev. 16, 143–163.

    Article  PubMed  CAS  Google Scholar 

  11. De Meyts P., Wallach B., Christoffersen, C. T., Ursø, B., Grønskov, K., Latus, L.-J., Yakushiji, F., Ilondo, M. M., and Shymko, R. M. (1994). Horm. Res. 42, 152–169.

    Article  PubMed  Google Scholar 

  12. Bornfeldt, K. E., Gidlf, R. A., Wasteson, A., Lake, M., Skottner, A., and Arnqvist, H. J. (1991). Diabetologia 34, 307–313.

    Article  PubMed  CAS  Google Scholar 

  13. Blum, W. F., Jenne, E. W., Reppin, F., Kietzmann, K., Ramke, M. B., and Bierich, J. R. (1989). Endocrinology 125: 766–772.

    Article  PubMed  CAS  Google Scholar 

  14. McCusker, R. H., Camacho-Hubner, C., Bayne, M. L., Cascieri, M. A., and Clemmon, D. R. (1990). J. Cell. Physiol. 144, 244–253.

    Article  PubMed  CAS  Google Scholar 

  15. Romagnolo, D., Akers, R. M., Byatt, J. C., Wong, E. A., and Turner, J. D. (1994). Endocr. J. 2, 375–384.

    CAS  Google Scholar 

  16. Huynh, H. T., Robitaille, G., and Turner, J. D. (1991). Exp. Cell. Res. 197, 191–199.

    Article  PubMed  CAS  Google Scholar 

  17. Zhao, X., McBride, B. W., Politis, I., Huynh, H. T., Akers, R. M., Burton, J. H., and Turner, J. D. (1992). J. Endocrinol. 134, 307–312.

    Article  PubMed  CAS  Google Scholar 

  18. Woodward, T. L., Akers, R. M., and Turner, J. D. (1994). Endocrine 2, 529–535.

    CAS  Google Scholar 

  19. McConnell, H. M., Owicki, J. C., Parce, J. W., Miller, D. L., Baxter, G. T., Wada, H. G., and Pitchford, S. (1992). Science 257, 1906–1912.

    Article  PubMed  CAS  Google Scholar 

  20. Baxter, G. T., Young, M.-L., Miller, D. L., and Owicki, J. C. (1994). Life Sci. 55, 573–583.

    Article  PubMed  CAS  Google Scholar 

  21. Chan, S. D. H., Antoniucci, D. M., Fok, K. S., Alajoki, M. L., Harkins, R. N., Thompson, S. A., and Wada, H. G., 1995). J Biol. Chem. 270, 22,608–22,613.

    Article  CAS  Google Scholar 

  22. Pitchford, S., De Moor, K., and Glaeser, B. S. (1995). Am. J. Physiol. C936-C943.

  23. Lauffenburger, D. A., Oerhtman, G.T., and Walker, L. (1998). Proc. Natl. Acad. Sci. USA 95, 15,368–15,373.

    Article  CAS  Google Scholar 

  24. Weber, M. S., Boyle, P. L., Corl, B. A., Wong, E. A.; Gwazdauskas, F. C., and Akers, R. M. (1988). Endocrine 8, 251–259.

    Article  Google Scholar 

  25. Romagnolo, D., Akers, R. M., Byatt, J. C., Wong, E. A., and Turner, J. D. (1994). Mol. Cell. Endocrinol. 102, 131–139.

    Article  CAS  Google Scholar 

  26. Oehrtman, G. T., Wiley, H. S., and Lauffenburger, D. A. (1998). Biotechnol. Bioeng. 57, 571–582.

    Article  PubMed  CAS  Google Scholar 

  27. Purup, S., Sejrsen, K., Foldager, J., and Akers, R. M. (1993), J. Endocrinol 139, 19–26.

    Article  PubMed  CAS  Google Scholar 

  28. Dickinson, K. E.J., Cohen, R. B., Skwish, S., Delaney, C. L., Serafino, R. P., Poss, M. A., Gu, Z., Ryono, D. E., Moreland, S., and Powell, J. R. (1994). Brit. J. Pharmacol. 113, 179–189.

    CAS  Google Scholar 

  29. Brown, M. J., Wood, M. D., Coldwell, M. C., and Bristow, D. R. (1997). Brit. J. Pharmacol. 121, 71–76.

    Article  CAS  Google Scholar 

  30. Chow, J. C., Condorelli, G., and Smith, R. J. (1998). J. Biol. chem. 273, 4672–4680.

    Article  PubMed  CAS  Google Scholar 

  31. Esposito, D. L., Blakesley, V. A., Koval, A. P., Scrimgeour, A. G., and LeRoith, D. (1997). Endocrinology 138, 2979–2988.

    Article  PubMed  CAS  Google Scholar 

  32. Kim, B., Cheng, H.-L.; Margolis, B., and Feldman, E. L. (1998). J. Biol. Chem. 273, 34,543–34,550.

    CAS  Google Scholar 

  33. Yau, L., Lukes, H., McDiarmid H., Werner, J., and Zahrakda, P. (1999). Eur. J. Biochem. 266, 1147–1157.

    Article  PubMed  CAS  Google Scholar 

  34. Duan, C., Liimatta, M. B., and Bottum, O. L. (1999). J. Biol. Chem. 274, 37,147–37,153.

    Article  CAS  Google Scholar 

  35. Laping, N. J., Olson, B. A., DeWolf, R. E., Albrightson, C. R., Fredrickson, T., King, C., Chirivella, M., Ziyadeh, F. N., and Nambi, P. (1998). Biochem. Pharmacol. 55, 227–234.

    Article  PubMed  CAS  Google Scholar 

  36. Hopkins, M. A., Rosser, M. P., Fernandes, P. B., et al. (1997). J. Neurosci. Methods 72, 167–174.

    Article  PubMed  CAS  Google Scholar 

  37. Twiss, J. L., Wada, H. G., Fok, K. S., Chan, S. D. H., Verity, A. N., Baxter, G. T., Shooter, E. M., and Sussman, H. H. (1998). J. Neurosci. Res. 51, 442–453.

    Article  PubMed  CAS  Google Scholar 

  38. Cohick, W. S. and Clemmons, D. R. (1993). Annu. Rev. Physiol. 55, 131–153.

    Article  PubMed  CAS  Google Scholar 

  39. Valentinis, B., Bhala, A., DeAngelis, T., Baserga, R., and Cohen, P. (1995). Mol. Endocrinol. 9, 361–367.

    Article  PubMed  CAS  Google Scholar 

  40. Rajah, R., Valentinis, B., and Cohen, P. (1997). J. Biol. Chem. 272, 12,181–12,188.

    Article  CAS  Google Scholar 

  41. Lauffenburger, D. A. and Linderman, J. J. (1993). Receptors: models for binding, trafficking, and signaling. Oxford University Press: New York.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Dairy Science, Virginia Polytechnic Institute and State University, Blacksburg, VA

    R. Michael Akers

  2. Department of Chemical Engineering, Virginia Polytechnic Institute and State University, 133 Randolph Hall, 24061, Blacksburg, VA

    Rose Marie Robinson & Kimberly E. Forsten

Authors
  1. Rose Marie Robinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R. Michael Akers
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kimberly E. Forsten
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kimberly E. Forsten.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Robinson, R.M., Akers, R.M. & Forsten, K.E. Real-time detection of insulin-like growth factor-1 stimulation of the MAC-T bovine mammary epithelial cell line. Endocr 13, 345–352 (2000). https://doi.org/10.1385/ENDO:13:3:345

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1385/ENDO:13:3:345

Key Words

Search

Navigation