, Volume 5, Issue 3, pp 341–349 | Cite as

Role of cell-cell adhesion in the regulation of prolactin gene expression by extracellular CaCl2

  • Melissa R. Lail-Trecker
  • Christopher J. Hanrahan
  • Bruce A. White
Original Articles


We have investigated a role for calcium-dependent cell-cell adhesion in the regulation of prolactin gene expression in rat pituitary GH3 cells. Cells cultured in a calcium-free, serum-free medium (SFM) express low levels of prolactin and growth hormone mRNA. As expected, addition of 0.5 mM CaCl2 to GH3 cells in SFM produced a specific, severalfold increase in prolactin mRNA levels. CaCl2 also promoted intercellular adhesion, during which cells assembled end-to-end in to cords. Prolactin mRNA increased after a delay of several hours. This latency period ranged from 4–12 h among different experiments, but always occurred after the onset of cell-cell adhesion. The voltage-sensitive calcium channel (VSCC) blocker, nitrendipine, inhibited the CaCl2-induced increase in prolactin mRNA without affecting cord formation. However, the VSCC agonist, BAY K-8644, was unable to induce prolactin gene expression prior to the onset of intercellular adhesion at 8 h, even though it produced a cellular response (tyrosine phosphorylation of a ca. 130-kDa protein) within 30 min. Blocking cell-cell adhesion inhibited the calcium-dependent induction of prolactin gene expression. Low levels (0.0025–0.02%) of trypsin blocked cell-cell adhesion and the prolactin mRNA induction by CaCl2 without affecting the levels of other mRNAs or cell-matrix adhesion. Heparin also specifically blocked the induction of both cell-cell adhesion and prolactin gene expression. Based on these data, we propose a role for both VSCCs and calcium-dependent cell-cell adhesion in the induction of prolactin gene expression by extracellular CaCl2.

Key Words

Calcium adhesion prolactin 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alcivar, A. A., Hake, L. E., Hardy, M. P., and Hecht, N. B. (1990).J. Biol. Chem. 265, 20,160–20,165.Google Scholar
  2. Bancroft, F. (1981). In:Functionally Differentiated Cell Lines. Sato, G. (ed.). Alan R. Liss: New York, pp. 47–59.Google Scholar
  3. Billis, W., Delidow, B., and White, B. (1992).Mol. Endocrinol. 6, 1277–1284.PubMedCrossRefGoogle Scholar
  4. Bradford, M. M. (1976).Anal. Biochem. 72, 248–254.PubMedCrossRefGoogle Scholar
  5. Calauti, E., Missero, C., Stein, P. L., Ezzell, R. M., and Dotto, G. P. (1995).Genes Dev. 9, 2279–2291.CrossRefGoogle Scholar
  6. Chronwell, B., Sands, S., Li, Z., and Shah, G. (1996).Endocrine 4, 27–33.CrossRefGoogle Scholar
  7. Cole, G. J. and Glaser, L. (1986).J. Cell Biol. 102, 403–412.PubMedCrossRefGoogle Scholar
  8. Day, R. and Maurer, R. (1990).Mol. Endocrinol. 4, 736–742.PubMedGoogle Scholar
  9. Delidow, B., Billis, W., Agarwal, P., and White, B. (1991).Mol. Endocrinol. 5, 1716–1722.PubMedGoogle Scholar
  10. Denef, C. (1985). In:Prolactin: Basic and Clinical Correlates. MacLeod, R., Thorner, M., and Scapagnini, U. (eds.), Liviana Press: Padua, pp. 53–57.Google Scholar
  11. Doherty, P., Ashton, S. V., Moore, S. E., and Walsh, F. S. (1991).Cell 67, 21–33.PubMedCrossRefGoogle Scholar
  12. Elenius, K. and Jalkanen, M. (1994).J. Cell Sci. 107, 2975–2982.PubMedGoogle Scholar
  13. Elias, K. A., Weiner, R. I., and Mellon, S. H. (1990).DNA Cell Biol. 9, 369–375.PubMedCrossRefGoogle Scholar
  14. Felix, R., Meza, U., and Cota, G. (1995).Endocrinology 136, 939–946.PubMedCrossRefGoogle Scholar
  15. Filvaroff, E., Stern, D. F., and Dotto, G. P. (1990).Mol. Cell. Biol. 10, 1164–1173.PubMedGoogle Scholar
  16. Fletcher, W., Anderson, N. J., and Everett, J. (1975).J. Cell Biol. 67, 469–476.PubMedCrossRefGoogle Scholar
  17. Frawley, L. and Boockfor, F. (1991).Endocr. Rev. 12, 337–355.PubMedGoogle Scholar
  18. Fujimori, T. and Takeichi, M. (1993).Mol. Biol. of the Cell. 4, 37–47.Google Scholar
  19. Greenberg, M. and Ziff, E. (1984).Nature 311, 433–438.PubMedCrossRefGoogle Scholar
  20. Guimond, S., Maccarana, M., Olwin, B. B., Lindahl, U., and Rapraeger, A. C. (1993).J. Biol. Chem. 268, 23,906–23,914.Google Scholar
  21. Harris, M. E., Bohni, R., Schneiderman, M. H., Ramamutrthy, L., Schumperli, D., and Marzluff, W. F. (1991).Mol. Cell. Biol. 11, 2416–2424.PubMedGoogle Scholar
  22. Hennings, H. and Holbrook, K. (1983).Exp. Cell Res. 143, 127–142.PubMedCrossRefGoogle Scholar
  23. Hennings, H., Michael, D., Cheng, C., Steinert, P., Holbrook, K., and Yuspa, S. (1980).Cell 19, 245–254.PubMedCrossRefGoogle Scholar
  24. Hinkle, P., Jackson, A., Thompson, T., Zavecki, A., Coppola, D., and Bancroft, C. (1988).Mol. Endocrinol. 2, 1132–1138.PubMedGoogle Scholar
  25. Hodivala, K. J. and Watt, F. M. (1994).J. Cell Biol. 124, 589–600.PubMedCrossRefGoogle Scholar
  26. Horvath, E. and Kovacs, K. (1988).J. Electron. Microsc. Tech. 8, 401–432.PubMedCrossRefGoogle Scholar
  27. Horvath, E., Kovacs, K., and Ezrin, C. (1977).IRCS Med. Sci. 5, 511–520.Google Scholar
  28. Ingraham, H., Albert, V., Chen, R., Crenshaw, E. I., Elsholtz, H., He, X., Kapiloff, M., Mangalam, H., Swanson, L., Treacy, M., and Rosenfeld, M. (1990).Annu. Rev. Physiol. 52, 773–791.PubMedCrossRefGoogle Scholar
  29. Jackson, A. and Bancroft, C. (1988).Mol. Endocrinol. 2, 1139–1144.PubMedGoogle Scholar
  30. Jue, S. F., Bradley, R. S., Rudnicki, J. A., Varmus, H. E., and Brown, A. M. (1992).Mol. Cell. Biol. 12, 321–328.PubMedGoogle Scholar
  31. Kan, M., Wang, F., Xu, J., Crabb, J. W., Hou, J., and McKeehan, W. L. (1993).Science 259, 1918–1921.PubMedCrossRefGoogle Scholar
  32. Kobayashi, H., Mizuki, T., Wada, A., and Izumi, F. (1992).Neuroscience 49, 437–441.PubMedCrossRefGoogle Scholar
  33. Laemmli, U. K. (1970).Nature 227, 680.PubMedCrossRefGoogle Scholar
  34. Lahr, G., Mayerhoffer, A., Bucher, S., Barthels, D., Wille, W., and Gratzl, M. (1993).Endocrinology 132, 1207–1217.PubMedCrossRefGoogle Scholar
  35. Maniatis, T., Fritsch, E., and Sambrook, J. (1982).Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, NY.Google Scholar
  36. Missale, C., Boroni, F., Sigala, S., Zanellato, A., Dal Toso, R., Balsari, A., and Spano, P. (1994).Endocrinology 135, 290–298.PubMedCrossRefGoogle Scholar
  37. Nakane, P. (1970).J. Histochem. Cytochem. 18, 9–21.PubMedGoogle Scholar
  38. Nelson, R. M., Ceccioni, O., Roberts, W. G., Aruffo, A., Linhardt, R. J., and Bevilacqua, M. P. (1993).Blood 82, 3253–3258.PubMedGoogle Scholar
  39. Nogami, H. and Yoshimura, F. (1982).Anat. Rec. 202, 261–274.PubMedCrossRefGoogle Scholar
  40. Preston, G., Billis, W., and White, B. (1990).Mol. Cell. Biol. 10, 442–448.PubMedGoogle Scholar
  41. Rapraeger, A. C. (1993).Curr. Opinion Cell Biol. 5, 844–853.PubMedCrossRefGoogle Scholar
  42. Reisfeld, S. and Vardimon, L. (1994).Mol. Endocrinol. 8, 1224–1233.PubMedCrossRefGoogle Scholar
  43. Rosenfeld, M. G., Nelson, C., Crenshaw, E. B., Elsholtz, H. P. A., Mangalam, H. J., Franxco, R., Waterman, M., Weinberger, C., Hollenberg, S. M., Giguere, V., Ong, E., and Evans, R. M. (1987).Recent Prog. Hormone Res. 43, 499–534.Google Scholar
  44. Saadat, S. and Thoenen, H. (1986).J. Cell Biol. 103, 1991–1997.PubMedCrossRefGoogle Scholar
  45. Tougard, C. and Tixier-Vidal, A. (1994). In:The Physiology of Reproduction, Second Edition. Knobil, E. and Neill, J. (eds.). Raven: New York, pp. 1711–1747.Google Scholar
  46. Volk, T., Volberg, T., Sabanay, I., and Geiger, B. (1990).Dev. Biol. 139, 314–326.PubMedCrossRefGoogle Scholar
  47. White, B. (1985).J. Biol. Chem. 260, 1213–1217.PubMedGoogle Scholar
  48. White, B. and Bancroft, C. (1987).Methods Enzymol. 139, 655–666.PubMedCrossRefGoogle Scholar
  49. White, B., Bauerle, L., and Bancroft, C. (1981).J. Biol. Chem. 256, 5942–5945.PubMedGoogle Scholar
  50. White, B., Power, E., and Fay, F. (1989).Mol. Endocrinol. 3, 1757–1764.PubMedGoogle Scholar
  51. Xanthopoulos, K. G., Mirkovitch, J., Decker, T., Kuo, C. F., and Darnell, J. E. (1989).Proc. Natl. Acad. Sci. USA 86, 4117–4121.PubMedCrossRefGoogle Scholar
  52. Yuspa, S., Kilkenney, A., Steinert, P., and Roop, D. (1989).J. Cell Biol. 109, 1207–1217.PubMedCrossRefGoogle Scholar
  53. Zhao, Y., Sudol, M., Hanafusa, H., and Krueger, J. (1992).Cell Biol. 89, 8298–8302.Google Scholar
  54. Zhao, Y., Uyttendaele, H., Krueger, J. G., Sudol, M., and Hanafusa, H. (1993).Mol. Cell. Biol. 13, 7507–7514.PubMedGoogle Scholar

Copyright information

© Humana Press Inc 1996

Authors and Affiliations

  • Melissa R. Lail-Trecker
    • 1
  • Christopher J. Hanrahan
    • 1
  • Bruce A. White
    • 1
  1. 1.Graduate Program in Developmental Biology, Department of AnatomyUniversity of Connecticut Health CenterFarmington

Personalised recommendations