Subcellular Localization of Components of the Dystrophin Glycoprotein Complex in Cultured Retinal Muller Glial Cells

Abstract

The dystrophin glycoprotein complex (DGC) is a membrane-associated protein complex binding extra-cellular matrix (ECM) molecules, such as laminin and forming a bridge towards the cytoskeleton. The molecular composition of the DGC is cell type dependent and it is involved in cell adhesion and motility. Here we present immunocytochemical localization of beta-dystroglycan, the central member of the DGC, utrophin and Dp71f the spliced 71 kDa dystrophin protein product of the DMD gene, in cultured retinal Muller glial cells. It is shown that beta-dystroglycan and utrophin are colocalized in clusters in all parts of Muller cells including the lamellipodium and leading edge of migrating cells. As a contrast, Dp71f labels are distinct from beta-dystroglycan and confined to the perinuclear cytoplasm of Muller cells indicating that Dp71f is not a member of the DGC in cultured Muller cells.

References

  1. 1.

    Aleman, V., Osorio, B., Chavez, O., Rendon, A., Momet, D., Martinez, D. (2001) Subcellular local-ization of Dp71 dystrophin isoforms in cultured hippocampal neurons and forebrain astrocytes. Histochem. Cell Biol. 115, 243–254.

    CAS  PubMed  Google Scholar 

  2. 2.

    Cavaldesi, M., Macchia, G., Barca, S., Defilippi, R., Tarone, G., Petrucci, T. C. (1999) Association of the dystroglycan complex isolated from bovine brain synaptosomes with proteins involved in signal transduction. J. Neurochem. 72, 1648–1655.

    CAS  Article  Google Scholar 

  3. 3.

    Claudepierre, T., Dalloz, C., Mornet, D., Matsumura, K., Sahel, J., Rendon, A. (2000) Characteriza-tion of the intermolecular associations of the dystrophin-associated glycoprotein complex in retinal Muller glial cells. J. Cell Sci. 113, 3409–3417.

    CAS  PubMed  Google Scholar 

  4. 4.

    Claudepierre, T., Mornet, D., Pannicke, T., Forster, V., Dalloz, C., Bolanos, F., Sahel, J., Reichen-bach, A., Rendon, A. (2000) Expression of Dp71 in Muller glial cells: a. comparison with utrophin-and dystrophin-associated proteins. Invest. Ophthalmol. Vis. Sci. 41, 294–304.

    CAS  PubMed  Google Scholar 

  5. 5.

    Claudepierre, T., Rodius, F., Frasson, M., Fontaine, V., Picaud, S., Dreyfus, H., Mornet, D., Rendon, A. (1999) Differential distribution of dystrophins in rat retina. Invest. Ophthalmol. Vis. Sci. 40, 1520–1529.

    CAS  PubMed  Google Scholar 

  6. 6.

    Durbeej, M., Campbell, K. R. (2002) Muscular dystrophies involving the dystrophin-glycoprotein complex: an overview of current mouse models. Curr. Opin. Genet. Dev. 12, 349–361.

    CAS  Article  Google Scholar 

  7. 7.

    Fabbrizio, E., Nudel, U., Hugon, G., Robert, A., Pons, F., Mornet, D. (1994) Characterization and localization of a. 77 kDa protein related to the dystrophin gene family. Biochem. J. 299, 359–365.

    CAS  Article  Google Scholar 

  8. 8.

    Garcia-Tovar, C. G., Luna, J., Mena, R., Soto-Zarate, C. I., Cortes, R., Perez, A., Leon-Avila, G., Mornet, D., Rendon, A., Hernandez, J. M. (2002) Dystrophin isoform Dp71 is present in lamellipo-dia and focal complexes in human astrocytoma cells U-373 MG. Acta Histochem. 104, 245–254.

    CAS  Article  Google Scholar 

  9. 9.

    Garcia-Tovar, C. G., Perez, A., Luna, J., Mena, R., Osorio, B., Aleman, V., Mondragon, R., Mornet, D., Rendon, A., Hernandez, J. M. (2001) Biochemical and histochemical analysis of 71 kDa dystrophin isoform (Dp71f) in rat brain. Acta Histochem. 103, 209–224.

    CAS  Article  Google Scholar 

  10. 10.

    Gorecki, D. C., Abdulrazzak, H., Lukasiuk, K., Barnard, E. A. (1997) Differential expression of syn-trophins and analysis of alternatively spliced dystrophin transcripts in the mouse brain. Eur. J. Neurosci. 9, 965–976.

    CAS  Article  Google Scholar 

  11. 11.

    Henry, M. D., Campbell, K. P. (1998) A. role for dystroglycan in basement membrane assembly. Cell 95, 859–870.

    CAS  Article  Google Scholar 

  12. 12.

    Hicks, D., Courtois, Y. (1999) The growth and behaviour of rat retinal Muller cells in vitro. An improved method for isolation and culture. Exp. Eye Res. 51, 119–129.

    Article  Google Scholar 

  13. 13.

    Howard, R. L., Klamut, H. J., Ray, R. N. (1998) Identification of a. novel actin binding site within the Dp71 dystrophin isoform. FEBS Letters 441, 337–341.

    CAS  Article  Google Scholar 

  14. 14.

    Ibraghimov-Beskrovnaya, O., Ervasti, J. M., Leveille, C. J., Slaughter, C. A., Sernett, S. W., Campbell, K. P. (1992) Primary structure of dystrophin-associated glycoproteins linking dystrophin to the extracellular matrix. Nature 355, 696–702.

    CAS  Article  Google Scholar 

  15. 15.

    Ilsley, J. L., Sudol, M., Winder, S. J. (2001) The interaction of dystrophin with beta-dystroglycan is regulated by tyrosine phosphorylation. Cell Signal. 13, 625–632.

    CAS  Article  Google Scholar 

  16. 16.

    Ilsley, J. L., Sudol, M., Winder, S. J. (2002) The WW domain: linking cell signalling to the membrane cytoskeleton. Cell Signal. 14, 183–189.

    CAS  Article  Google Scholar 

  17. 17.

    James, M., Nuttall, A., Ilsley, J. L., Ottersbach, K., Tinsley, J. M., Sudol, M., Winder, S. J. (2000) Adhesion-dependent tyrosine phosphorylation of (beta)-dystroglycan regulates its interaction with utrophin. J. Cell Sci. 113, 1717–1726.

    CAS  PubMed  Google Scholar 

  18. 18.

    Leibovitz, S., Meshorer, A., Fridman, Y., Wieneke, S., Jockusch, H., Yaffe, D., Nudel, U. (2002) Exogenous Dp71 is a. dominant negative competitor of dystrophin in skeletal muscle. Neuromuscul. Disord. 12, 836–844.

    Article  Google Scholar 

  19. 19.

    Lumeng, C., Phelps, S., Crawford, G. E., Walden, R. D., Barald, K., Chamberlain J. S. (1999) Interactions between beta 2-syntrophin and a. family of microtubule-associated serine/threonine kinases. Nat. Neurosci. 27, 611–617.

    Article  Google Scholar 

  20. 20.

    Méhes, E., Czirok, A., Hegedüs, B., Vicsek, T., Jancsik, V. (2002) Laminin-1 increases motility, path-searching, and process dynamism of rat and mouse Muller glial cells in vitro: implication of rela-tionship between cell behavior and formation of retinal morphology. Cell Motil. Cytoskeleton 53, 203–213.

    Article  Google Scholar 

  21. 21.

    Pereboev, A. V., Ahmed, N., thi Man, N., Morris, G. E. (2001) Epitopes in the interacting regions of beta-dystroglycan (PPxY motif) and dystrophin (WW domain). Biochim. Biophys. Acta 1527, 54–60.

    CAS  Article  Google Scholar 

  22. 22.

    Pons, F., Robert, A., Fabbrizio, E., Hugon, G., Califano, J. P. C., Fehrentz, J. A., Martinez, J., Mornet, D. (1994) Utrophin localization in normal and dystrophin-deficient heart. Circulation 90, 369–374.

    CAS  Article  Google Scholar 

  23. 23.

    Rentschler, S., Linn, H., Deininger, K., Bedford, M. T., Espanel, X., Sudol, M. (1999) The WW domain of dystrophin requires EF-hands region to interact with beta-dystroglycan. Biol. Chem. 380, 431–142.

    CAS  Article  Google Scholar 

  24. 24.

    Sotgia, E., Lee, H., Bedford, M. T., Petrucci, T., Sudol, M., Lisanti, M. P. (2001) Tyrosine phospho-rylation of beta-dystroglycan at its WW domain binding motif, PPxY, recruits SH2 domain contain-ing proteins. Biochemistry 40, 14585–14592.

    CAS  Article  Google Scholar 

  25. 25.

    Winder, S. J. (2001) The complexities of dystroglycan. Trends Biochem. Sci. 26, 118–124.

    CAS  Article  Google Scholar 

Download references

Acknowledgement

This work was supported by the Hungarian Scientific Research Fund (OTKA, T 037597) for V. J.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Veronika Jancsik.

Rights and permissions

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Reprints and Permissions

About this article

Cite this article

Méhes, E., Mornet, D. & Jancsik, V. Subcellular Localization of Components of the Dystrophin Glycoprotein Complex in Cultured Retinal Muller Glial Cells. BIOLOGIA FUTURA 54, 241–252 (2003). https://doi.org/10.1556/ABiol.54.2003.3-4.3

Download citation

Keywords

  • Immunocytochemistry
  • rat
  • dystrophin
  • utrophin
  • cytoskeleton