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. 1984 Apr 1;98(4):1565–1571. doi: 10.1083/jcb.98.4.1565

Role of calcium and calmodulin in hemidesmosome formation in vitro

PMCID: PMC2113221  PMID: 6715411

Abstract

Intact epithelial sheets were removed from rabbit corneas using Dispase II, a bacterial neutral protease. The freed sheets were placed on denuded corneal basal laminae and incubated at 35 degrees C for 3, 6, 18, or 24 h. Epithelial-basal lamina preparations were incubated in culture medium that either contained (a) varying concentrations of Ca2+ ions, (b) calmodulin antagonists, (c) exogenous calmodulin following an initial 6-h incubation in the presence of antagonists, or that lacked (d) Mg2+ ions. Tissues were processed for electron microscopy, and micrographs were taken of basal cell membranes. At least four experiments were conducted for each treatment, and for each experiment the total number of hemidesmosomes were counted along the basal membrane-basal lamina surface of eight cells. The number of hemidesmosomes formed was directly proportional to the increasing concentration of Ca2+. The presence of absence of Mg2+ ions did not change the numbers of hemidesmosomes formed. Calmodulin antagonists inhibited hemidesmosome formation, and this inhibition was reversed by the addition of calmodulin. Thus, hemidesmosome formation is Ca2+ dependent and appears to be mediated by a calmodulin-regulated mechanism.

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Selected References

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  1. Anderson R. A. Actin filaments in normal and migrating corneal epithelial cells. Invest Ophthalmol Vis Sci. 1977 Feb;16(2):161–166. [PubMed] [Google Scholar]
  2. Beerens E. G., Slot J. W., van der Leun J. C. Rapid regeneration of the dermal-epidermal junction after partial separation by vacuum: an electron microscopic study. J Invest Dermatol. 1975 Dec;65(6):513–521. doi: 10.1111/1523-1747.ep12610200. [DOI] [PubMed] [Google Scholar]
  3. Boynton A. L., Whitfield J. F., MacManus J. P. Calmodulin stimulates DNA synthesis by rat liver cells. Biochem Biophys Res Commun. 1980 Jul 31;95(2):745–749. doi: 10.1016/0006-291x(80)90849-9. [DOI] [PubMed] [Google Scholar]
  4. Brackenbury R., Rutishauser U., Edelman G. M. Distinct calcium-independent and calcium-dependent adhesion systems of chicken embryo cells. Proc Natl Acad Sci U S A. 1981 Jan;78(1):387–391. doi: 10.1073/pnas.78.1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Buck R. C. Cell migration in repair of mouse corneal epithelium. Invest Ophthalmol Vis Sci. 1979 Aug;18(8):767–784. [PubMed] [Google Scholar]
  7. Cheung W. Y. Cyclic 3',5'-nucleotide phosphodiesterase. Demonstration of an activator. Biochem Biophys Res Commun. 1970 Feb 6;38(3):533–538. doi: 10.1016/0006-291x(70)90747-3. [DOI] [PubMed] [Google Scholar]
  8. Connor C. G., Brady R. C., Brownstein B. L. Trifluoperazine inhibits spreading and migration of cells in culture. J Cell Physiol. 1981 Sep;108(3):299–307. doi: 10.1002/jcp.1041080303. [DOI] [PubMed] [Google Scholar]
  9. Gipson I. K., Grill S. M. A technique for obtaining sheets of intact rabbit corneal epithelium. Invest Ophthalmol Vis Sci. 1982 Aug;23(2):269–273. [PubMed] [Google Scholar]
  10. Gipson I. K., Kiorpes T. C. Epithelial sheet movement: protein and glycoprotein synthesis. Dev Biol. 1982 Jul;92(1):259–262. doi: 10.1016/0012-1606(82)90170-1. [DOI] [PubMed] [Google Scholar]
  11. Grunwald G. B., Geller R. L., Lilien J. Enzymatic dissection of embryonic cell adhesive mechanisms. J Cell Biol. 1980 Jun;85(3):766–776. doi: 10.1083/jcb.85.3.766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hennings H., Holbrook K. A. Calcium regulation of cell-cell contact and differentiation of epidermal cells in culture. An ultrastructural study. Exp Cell Res. 1983 Jan;143(1):127–142. doi: 10.1016/0014-4827(83)90115-5. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Hidaka H., Asano M., Tanaka T. Activity-structure relationship of calmodulin antagonists, Naphthalenesulfonamide derivatives. Mol Pharmacol. 1981 Nov;20(3):571–578. [PubMed] [Google Scholar]
  15. Jones J. C., Goldman A. E., Steinert P. M., Yuspa S., Goldman R. D. Dynamic aspects of the supramolecular organization of intermediate filament networks in cultured epidermal cells. Cell Motil. 1982;2(3):197–213. doi: 10.1002/cm.970020302. [DOI] [PubMed] [Google Scholar]
  16. Kakiuchi S., Yamazaki R. Calcium dependent phosphodiesterase activity and its activating factor (PAF) from brain studies on cyclic 3',5'-nucleotide phosphodiesterase (3). Biochem Biophys Res Commun. 1970 Dec 9;41(5):1104–1110. doi: 10.1016/0006-291x(70)90199-3. [DOI] [PubMed] [Google Scholar]
  17. Krawczyk W. S., Wilgram G. F. Hemidesmosome and desmosome morphogenesis during epidermal wound healing. J Ultrastruct Res. 1973 Oct;45(1):93–101. doi: 10.1016/s0022-5320(73)90035-x. [DOI] [PubMed] [Google Scholar]
  18. LaPorte D. C., Wierman B. M., Storm D. R. Calcium-induced exposure of a hydrophobic surface on calmodulin. Biochemistry. 1980 Aug 5;19(16):3814–3819. doi: 10.1021/bi00557a025. [DOI] [PubMed] [Google Scholar]
  19. Levin R. M., Weiss B. Binding of trifluoperazine to the calcium-dependent activator of cyclic nucleotide phosphodiesterase. Mol Pharmacol. 1977 Jul;13(4):690–697. [PubMed] [Google Scholar]
  20. Lew P. D., Stossel T. P. Calcium transport by macrophage plasma membranes. J Biol Chem. 1980 Jun 25;255(12):5841–5846. [PubMed] [Google Scholar]
  21. Means A. R., Chafouleas J. G. Regulation by and of calmodulin in mammalian cells. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):903–908. doi: 10.1101/sqb.1982.046.01.084. [DOI] [PubMed] [Google Scholar]
  22. Pitelka D. R., Taggart B. N., Hamamoto S. T. Effects of extracellular calcium depletion on membrane topography and occluding junctions of mammary epithelial cells in culture. J Cell Biol. 1983 Mar;96(3):613–624. doi: 10.1083/jcb.96.3.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tanaka T., Hidaka H. Hydrophobic regions function in calmodulin-enzyme(s) interactions. J Biol Chem. 1980 Dec 10;255(23):11078–11080. [PubMed] [Google Scholar]
  24. Tanaka T., Ohmura T., Hidaka H. Hydrophobic interaction of the Ca2+-calmodulin complex with calmodulin antagonists. Naphthalenesulfonamide derivatives. Mol Pharmacol. 1982 Sep;22(2):403–407. [PubMed] [Google Scholar]
  25. Thomas W. A., Edelman B. A., Lobel S. M., Breitbart A. S., Steinberg M. S. Two chick embryonic adhesion systems: molecular vs tissue specificity. J Supramol Struct Cell Biochem. 1981;16(1):15–27. doi: 10.1002/jsscb.1981.380160103. [DOI] [PubMed] [Google Scholar]
  26. Wallace R. W., Tallant E. A., Cheung W. Y. Multifunctional role of calmodulin in biologic processes. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):893–901. doi: 10.1101/sqb.1982.046.01.083. [DOI] [PubMed] [Google Scholar]
  27. Weiss B., Levin R. M. Mechanism for selectively inhibiting the activation of cyclic nucleotide phosphodiesterase and adenylate cyclase by antipsychotic agents. Adv Cyclic Nucleotide Res. 1978;9:285–303. [PubMed] [Google Scholar]
  28. Weiss B., Prozialeck W. C., Wallace T. L. Interaction of drugs with calmodulin. Biochemical, pharmacological and clinical implications. Biochem Pharmacol. 1982 Jul 1;31(13):2217–2226. doi: 10.1016/0006-2952(82)90104-6. [DOI] [PubMed] [Google Scholar]

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