Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1991 Aug 1;114(3):545–555. doi: 10.1083/jcb.114.3.545

Regulation of connexin 43-mediated gap junctional intercellular communication by Ca2+ in mouse epidermal cells is controlled by E- cadherin

PMCID: PMC2289094  PMID: 1650371

Abstract

Gap junctional intercellular communication (GJIC) of cultured mouse epidermal cells is mediated by a gap junction protein, connexin 43, and is dependent on the calcium concentration in the medium, with higher GJIC in a high-calcium (1.2 mM) medium. In several mouse epidermal cell lines, we found a good correlation between the level of GJIC and that of immunohistochemical staining of E-cadherin, a calcium-dependent cell adhesion molecule, at cell-cell contact areas. The variant cell line P3/22 showed both low GJIC and E-cadherin protein expression in low- and high-Ca2+ media. P3/22 cells showed very low E-cadherin mRNA expression. To test directly whether E-cadherin is involved in the Ca(2+)-dependent regulation of GJIC, we transfected the E-cadherin expression vector into P3/22 cells and obtained several stable clones which expressed high levels of E-cadherin mRNA. All transfectants expressed E-cadherin molecules at cell-cell contact areas in a calcium- dependent manner. GJIC was also observed in these transfectants and was calcium dependent. These results suggest that Ca(2+)-dependent regulation of GJIC in mouse epidermal cells is directly controlled by a calcium-dependent cell adhesion molecule, E-cadherin. Furthermore, several lines of evidence suggest that GJIC control by E-cadherin involves posttranslational regulation (assembly and/or function) of the gap junction protein connexin 43.

Full Text

The Full Text of this article is available as a PDF (2.8 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beyer E. C., Kistler J., Paul D. L., Goodenough D. A. Antisera directed against connexin43 peptides react with a 43-kD protein localized to gap junctions in myocardium and other tissues. J Cell Biol. 1989 Feb;108(2):595–605. doi: 10.1083/jcb.108.2.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beyer E. C., Paul D. L., Goodenough D. A. Connexin43: a protein from rat heart homologous to a gap junction protein from liver. J Cell Biol. 1987 Dec;105(6 Pt 1):2621–2629. doi: 10.1083/jcb.105.6.2621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brackenbury R., Greenberg M. E., Edelman G. M. Phenotypic changes and loss of N-CAM-mediated adhesion in transformed embryonic chicken retinal cells. J Cell Biol. 1984 Dec;99(6):1944–1954. doi: 10.1083/jcb.99.6.1944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brennan J. K., Mansky J., Roberts G., Lichtman M. A. Improved methods for reducing calcium and magnesium concentrations in tissue culture medium: application to studies of lymphoblast proliferation in vitro. In Vitro. 1975 Nov-Dec;11(6):354–360. doi: 10.1007/BF02616371. [DOI] [PubMed] [Google Scholar]
  5. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  6. Davidson J. S., Baumgarten I. M., Harley E. H. Effects of extracellular calcium and magnesium on junctional intercellular communication in human fibroblasts. Exp Cell Res. 1984 Dec;155(2):406–412. doi: 10.1016/0014-4827(84)90201-5. [DOI] [PubMed] [Google Scholar]
  7. Dotto G. P., el-Fouly M. H., Nelson C., Trosko J. E. Similar and synergistic inhibition of gap-junctional communication by ras transformation and tumor promoter treatment of mouse primary keratinocytes. Oncogene. 1989 May;4(5):637–641. [PubMed] [Google Scholar]
  8. Edelman G. M. Cell adhesion molecules in the regulation of animal form and tissue pattern. Annu Rev Cell Biol. 1986;2:81–116. doi: 10.1146/annurev.cb.02.110186.000501. [DOI] [PubMed] [Google Scholar]
  9. Eidelman S., Damsky C. H., Wheelock M. J., Damjanov I. Expression of the cell-cell adhesion glycoprotein cell-CAM 120/80 in normal human tissues and tumors. Am J Pathol. 1989 Jul;135(1):101–110. [PMC free article] [PubMed] [Google Scholar]
  10. Enomoto T., Yamasaki H. Lack of intercellular communication between chemically transformed and surrounding nontransformed BALB/c 3T3 cells. Cancer Res. 1984 Nov;44(11):5200–5203. [PubMed] [Google Scholar]
  11. Fearon E. R., Cho K. R., Nigro J. M., Kern S. E., Simons J. W., Ruppert J. M., Hamilton S. R., Preisinger A. C., Thomas G., Kinzler K. W. Identification of a chromosome 18q gene that is altered in colorectal cancers. Science. 1990 Jan 5;247(4938):49–56. doi: 10.1126/science.2294591. [DOI] [PubMed] [Google Scholar]
  12. Finbow M., Yancey S. B., Johnson R., Revel J. P. Independent lines of evidence suggesting a major gap junctional protein with a molecular weight of 26,000. Proc Natl Acad Sci U S A. 1980 Feb;77(2):970–974. doi: 10.1073/pnas.77.2.970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Frixen U. H., Behrens J., Sachs M., Eberle G., Voss B., Warda A., Löchner D., Birchmeier W. E-cadherin-mediated cell-cell adhesion prevents invasiveness of human carcinoma cells. J Cell Biol. 1991 Apr;113(1):173–185. doi: 10.1083/jcb.113.1.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Goodenough D. A., Paul D. L., Jesaitis L. Topological distribution of two connexin32 antigenic sites in intact and split rodent hepatocyte gap junctions. J Cell Biol. 1988 Nov;107(5):1817–1824. doi: 10.1083/jcb.107.5.1817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Greenberg M. E., Brackenbury R., Edelman G. M. Alteration of neural cell adhesion molecule (N-CAM) expression after neuronal cell transformation by Rous sarcoma virus. Proc Natl Acad Sci U S A. 1984 Feb;81(3):969–973. doi: 10.1073/pnas.81.3.969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hamel E., Katoh F., Mueller G., Birchmeier W., Yamasaki H. Transforming growth factor beta as a potent promoter in two-stage BALB/c 3T3 cell transformation. Cancer Res. 1988 May 15;48(10):2832–2836. [PubMed] [Google Scholar]
  17. Hashimoto M., Niwa O., Nitta Y., Takeichi M., Yokoro K. Unstable expression of E-cadherin adhesion molecules in metastatic ovarian tumor cells. Jpn J Cancer Res. 1989 May;80(5):459–463. doi: 10.1111/j.1349-7006.1989.tb02336.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hennings H., Michael D., Cheng C., Steinert P., Holbrook K., Yuspa S. H. Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell. 1980 Jan;19(1):245–254. doi: 10.1016/0092-8674(80)90406-7. [DOI] [PubMed] [Google Scholar]
  19. Johnson G. D., Nogueira Araujo G. M. A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods. 1981;43(3):349–350. doi: 10.1016/0022-1759(81)90183-6. [DOI] [PubMed] [Google Scholar]
  20. Kam E., Melville L., Pitts J. D. Patterns of junctional communication in skin. J Invest Dermatol. 1986 Dec;87(6):748–753. doi: 10.1111/1523-1747.ep12456937. [DOI] [PubMed] [Google Scholar]
  21. Kanno Y., Sasaki Y., Shiba Y., Yoshida-Noro C., Takeichi M. Monoclonal antibody ECCD-1 inhibits intercellular communication in teratocarcinoma PCC3 cells. Exp Cell Res. 1984 May;152(1):270–274. doi: 10.1016/0014-4827(84)90253-2. [DOI] [PubMed] [Google Scholar]
  22. Kawamura H., Strickland J. E., Yuspa S. H. Association of resistance to terminal differentiation with initiation of carcinogenesis in adult mouse epidermal cells. Cancer Res. 1985 Jun;45(6):2748–2752. [PubMed] [Google Scholar]
  23. Keane R. W., Mehta P. P., Rose B., Honig L. S., Loewenstein W. R., Rutishauser U. Neural differentiation, NCAM-mediated adhesion, and gap junctional communication in neuroectoderm. A study in vitro. J Cell Biol. 1988 Apr;106(4):1307–1319. doi: 10.1083/jcb.106.4.1307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Klann R. C., Fitzgerald D. J., Piccoli C., Slaga T. J., Yamasaki H. Gap-junctional intercellular communication in epidermal cell lines from selected stages of SENCAR mouse skin carcinogenesis. Cancer Res. 1989 Feb 1;49(3):699–705. [PubMed] [Google Scholar]
  25. Kumar N. M., Gilula N. B. Cloning and characterization of human and rat liver cDNAs coding for a gap junction protein. J Cell Biol. 1986 Sep;103(3):767–776. doi: 10.1083/jcb.103.3.767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lo C. W., Gilula N. B. Gap junctional communication in the post-implantation mouse embryo. Cell. 1979 Oct;18(2):411–422. doi: 10.1016/0092-8674(79)90060-6. [DOI] [PubMed] [Google Scholar]
  27. Lo C. W., Gilula N. B. Gap junctional communication in the preimplantation mouse embryo. Cell. 1979 Oct;18(2):399–409. doi: 10.1016/0092-8674(79)90059-x. [DOI] [PubMed] [Google Scholar]
  28. Loewenstein W. R. Junctional intercellular communication and the control of growth. Biochim Biophys Acta. 1979 Feb 4;560(1):1–65. doi: 10.1016/0304-419x(79)90002-7. [DOI] [PubMed] [Google Scholar]
  29. Loewenstein W. R. The cell-to-cell channel of gap junctions. Cell. 1987 Mar 13;48(5):725–726. doi: 10.1016/0092-8674(87)90067-5. [DOI] [PubMed] [Google Scholar]
  30. Mege R. M., Matsuzaki F., Gallin W. J., Goldberg J. I., Cunningham B. A., Edelman G. M. Construction of epithelioid sheets by transfection of mouse sarcoma cells with cDNAs for chicken cell adhesion molecules. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7274–7278. doi: 10.1073/pnas.85.19.7274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Menon G. K., Grayson S., Elias P. M. Ionic calcium reservoirs in mammalian epidermis: ultrastructural localization by ion-capture cytochemistry. J Invest Dermatol. 1985 Jun;84(6):508–512. doi: 10.1111/1523-1747.ep12273485. [DOI] [PubMed] [Google Scholar]
  32. Mesnil M., Yamasaki H. Selective gap-junctional communication capacity of transformed and non-transformed rat liver epithelial cell lines. Carcinogenesis. 1988 Aug;9(8):1499–1502. doi: 10.1093/carcin/9.8.1499. [DOI] [PubMed] [Google Scholar]
  33. Meyer D. J., Yancey S. B., Revel J. P. Intercellular communication in normal and regenerating rat liver: a quantitative analysis. J Cell Biol. 1981 Nov;91(2 Pt 1):505–523. doi: 10.1083/jcb.91.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Milks L. C., Kumar N. M., Houghten R., Unwin N., Gilula N. B. Topology of the 32-kd liver gap junction protein determined by site-directed antibody localizations. EMBO J. 1988 Oct;7(10):2967–2975. doi: 10.1002/j.1460-2075.1988.tb03159.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Miller D. R., Viaje A., Aldaz C. M., Conti C. J., Slaga T. J. Terminal differentiation-resistant epidermal cells in mice undergoing two-stage carcinogenesis. Cancer Res. 1987 Apr 1;47(7):1935–1940. [PubMed] [Google Scholar]
  36. Nagafuchi A., Takeichi M. Cell binding function of E-cadherin is regulated by the cytoplasmic domain. EMBO J. 1988 Dec 1;7(12):3679–3684. doi: 10.1002/j.1460-2075.1988.tb03249.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Nose A., Nagafuchi A., Takeichi M. Expressed recombinant cadherins mediate cell sorting in model systems. Cell. 1988 Sep 23;54(7):993–1001. doi: 10.1016/0092-8674(88)90114-6. [DOI] [PubMed] [Google Scholar]
  38. Nose A., Nagafuchi A., Takeichi M. Isolation of placental cadherin cDNA: identification of a novel gene family of cell-cell adhesion molecules. EMBO J. 1987 Dec 1;6(12):3655–3661. doi: 10.1002/j.1460-2075.1987.tb02698.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Odin P., Obrink B. Dynamic expression of the cell adhesion molecule cell-CAM 105 in fetal and regenerating rat liver. Exp Cell Res. 1986 May;164(1):103–114. doi: 10.1016/0014-4827(86)90458-1. [DOI] [PubMed] [Google Scholar]
  40. Ozawa M., Engel J., Kemler R. Single amino acid substitutions in one Ca2+ binding site of uvomorulin abolish the adhesive function. Cell. 1990 Nov 30;63(5):1033–1038. doi: 10.1016/0092-8674(90)90506-a. [DOI] [PubMed] [Google Scholar]
  41. Ozawa M., Ringwald M., Kemler R. Uvomorulin-catenin complex formation is regulated by a specific domain in the cytoplasmic region of the cell adhesion molecule. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4246–4250. doi: 10.1073/pnas.87.11.4246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Paul D. L. Molecular cloning of cDNA for rat liver gap junction protein. J Cell Biol. 1986 Jul;103(1):123–134. doi: 10.1083/jcb.103.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Pitts J. D., Finbow M. E. The gap junction. J Cell Sci Suppl. 1986;4:239–266. doi: 10.1242/jcs.1986.supplement_4.15. [DOI] [PubMed] [Google Scholar]
  44. Shimoyama Y., Hirohashi S., Hirano S., Noguchi M., Shimosato Y., Takeichi M., Abe O. Cadherin cell-adhesion molecules in human epithelial tissues and carcinomas. Cancer Res. 1989 Apr 15;49(8):2128–2133. [PubMed] [Google Scholar]
  45. Shirayoshi Y., Nose A., Iwasaki K., Takeichi M. N-linked oligosaccharides are not involved in the function of a cell-cell binding glycoprotein E-cadherin. Cell Struct Funct. 1986 Sep;11(3):245–252. doi: 10.1247/csf.11.245. [DOI] [PubMed] [Google Scholar]
  46. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  47. Spray D. C., Fujita M., Saez J. C., Choi H., Watanabe T., Hertzberg E., Rosenberg L. C., Reid L. M. Proteoglycans and glycosaminoglycans induce gap junction synthesis and function in primary liver cultures. J Cell Biol. 1987 Jul;105(1):541–551. doi: 10.1083/jcb.105.1.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Traub O., Look J., Dermietzel R., Brümmer F., Hülser D., Willecke K. Comparative characterization of the 21-kD and 26-kD gap junction proteins in murine liver and cultured hepatocytes. J Cell Biol. 1989 Mar;108(3):1039–1051. doi: 10.1083/jcb.108.3.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Yamasaki H., Hollstein M., Mesnil M., Martel N., Aguelon A. M. Selective lack of intercellular communication between transformed and nontransformed cells as a common property of chemical and oncogene transformation of BALB/c 3T3 cells. Cancer Res. 1987 Nov 1;47(21):5658–5664. [PubMed] [Google Scholar]
  50. Yamasaki H., Huberman E., Sachs L. Regulation of aryl hydrocarbon (benzo-(A)-pyrene) hydroxylase activity in mammalian cells. Induction of hydroxylase activity by N6,O2'-dibutyryl8 adenosine 3':5'-monophosphate and aminophylline. J Biol Chem. 1975 Oct 10;250(19):7766–7770. [PubMed] [Google Scholar]
  51. Yancey S. B., John S. A., Lal R., Austin B. J., Revel J. P. The 43-kD polypeptide of heart gap junctions: immunolocalization, topology, and functional domains. J Cell Biol. 1989 Jun;108(6):2241–2254. doi: 10.1083/jcb.108.6.2241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Yoshida-Noro C., Suzuki N., Takeichi M. Molecular nature of the calcium-dependent cell-cell adhesion system in mouse teratocarcinoma and embryonic cells studied with a monoclonal antibody. Dev Biol. 1984 Jan;101(1):19–27. doi: 10.1016/0012-1606(84)90112-x. [DOI] [PubMed] [Google Scholar]
  53. Yuspa S. H., Morgan D. L. Mouse skin cells resistant to terminal differentiation associated with initiation of carcinogenesis. Nature. 1981 Sep 3;293(5827):72–74. doi: 10.1038/293072a0. [DOI] [PubMed] [Google Scholar]
  54. Zhang J. T., Nicholson B. J. Sequence and tissue distribution of a second protein of hepatic gap junctions, Cx26, as deduced from its cDNA. J Cell Biol. 1989 Dec;109(6 Pt 2):3391–3401. doi: 10.1083/jcb.109.6.3391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. el Aoumari A., Fromaget C., Dupont E., Reggio H., Durbec P., Briand J. P., Böller K., Kreitman B., Gros D. Conservation of a cytoplasmic carboxy-terminal domain of connexin 43, a gap junctional protein, in mammal heart and brain. J Membr Biol. 1990 May;115(3):229–240. doi: 10.1007/BF01868638. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES