Molecular & Cellular Toxicology

The Korean Society of Toxicogenomics and Toxicoproteomics (대한독성유전 단백체학회)
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An Ototoxic Antibiotic Gentamicin Can Increase PKA-caveolin-1 Signaling Pathway in Differentiated Vestibular Cell Line (UB/UE-1)

  • Kim, Kyu-Sung (Department of Otolaryngology-Head and Neck Surgery College of Medicine Inha University) ;
  • Cho, Byung-Han (Department of Otolaryngology-Head and Neck Surgery College of Medicine Inha University) ;
  • Choi, Ho-Seok (Department of Otolaryngology-Head and Neck Surgery College of Medicine Inha University) ;
  • Park, Chang-Shin (Department of Pharmacology and Medicinal Toxicology Research Center, Inha University) ;
  • Jung, Yoon-Gun (Department of Otolaryngology-Head and Neck Surgery College of Medicine Inha University) ;
  • Kim, Young-Mo (Department of Otolaryngology-Head and Neck Surgery College of Medicine Inha University) ;
  • Jang, Tae-Young (Department of Otolaryngology-Head and Neck Surgery College of Medicine Inha University)
  • Published : 2008.09.30
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Abstract

Caveolin proteins are mediators of cell death or the survival of injured cells, and they are inhibitors of various signaling pathways. The expression of caveolin-, which is involved in the protein kinase A (PKA) signaling pathway, was examined in the differentiated mouse vestibular cell line UB/UE-1 after gentamicin ototoxicity. Caveolae in the vestibular hair cell of healthy guinea pigs were observed through an electron microscope. UB/UE-1 cells were cultured at 95% $CO_2$ with 5% $O_2$ at $33^{\circ}C$ for 48 hours and at 95% $CO_2$ with 5% $O_2$ at $39^{\circ}C$ for 24 hours for differentiation. Cells were treated with 1 mM gentamicin, 0.02 mM H89 (PKA inhibitor), and then incubated for 24 hours. Caveolin-1 expression was examined by western blotting and PKA activity by a $PepTag^{(R)}$ assay. Caveolae were observed in the vestibular hair cells of healthy guinea pigs by electron microscopy. Caveolin-1 was expressed spontaneously in differentiated UB/UE-1 cells and increased after gentamicin treatment. PKA was also over-activated by gentamicin treatment. Both gentamicin-induced caveolin-1 expression and PKA over-activation were inhibited by H89. These results indicate that gentamicin-induced caveolin-1 expression is mediated by the PKA signaling pathway. We conclude that caveolae/ caveolin activity, induced via a PKA signaling pathway, may be one of the mechanisms of gentamicin-induced ototoxicity.

Keywords

References

  1. Frank, P. G. & Lisanti, M. P. Caveolin-1 and caveolae in atherosclerosis: differential roles in fatty streak formation and neointimal hyperplasia. Curr Opin Lipidol 15:523-529 (2004) https://doi.org/10.1097/00041433-200410000-00005
  2. Razani, B., Woodman, S. E. & Lisanti, M. P. Caveolae: from cell biology to animal physiology. Pharmacol Rev 54:431-467 (2002) https://doi.org/10.1124/pr.54.3.431
  3. Fra, A. M., Williamson, E., Simons, K. & Parton, R. G. De novo formation of caveolae in lymphocytes by expression of VIP21-caveolin. Proc Natl Acad Sci U S A 92:8655-8659 (1995)
  4. Engelman, J. A. et al. Recombinant expression of caveolin-1 in oncogenically transformed cells abrogates anchorage-independent growth. J Biol Chem 272:16374-16381 (1997) https://doi.org/10.1074/jbc.272.26.16374
  5. Li, S. et al. Mutational analysis of caveolin-induced vesicle formation. Expression of caveolin-1 recruits caveolin-2 to caveolae membranes. FEBS Lett 434: 127-134 (1998) https://doi.org/10.1016/S0014-5793(98)00945-4
  6. Galbiati, F. et al. Targeted downregulation of caveolin- 1 is sufficient to drive cell transformation and hyperactivate the p42/44 MAP kinase cascade. EMBO J 17:6633-6648 (1998) https://doi.org/10.1093/emboj/17.22.6633
  7. Liu, J., Lee, P., Galbiati, F., Kitsis, R. N. & Lisanti, M. P. Caveolin-1 expression sensitizes fibroblastic and epithelial cells to apoptotic stimulation. Am J Physiol Cell Physiol 280:C823-835 (2001) https://doi.org/10.1152/ajpcell.2001.280.4.C823
  8. Simionescu, N., Siminoescu, M. & Palade, G. E. Permeability of muscle capillaries to small heme-peptides. Evidence for the existence of patent transendothelial channels. J Cell Biol 64:586-607 (1975) https://doi.org/10.1083/jcb.64.3.586
  9. Takei, K. & Haucke, V. Clathrin-mediated endocytosis: membrane factors pull the trigger. Trends Cell Biol 11:385-391 (2001) https://doi.org/10.1016/S0962-8924(01)02082-7
  10. Gumbleton, M. Caveolae as potential macromolecule trafficking compartments within alveolar epithelium. Adv Drug Deliv Rev 49:281-300 (2001) https://doi.org/10.1016/S0169-409X(01)00142-9
  11. Montesano, R., Roth, J., Robert, A. & Orci, L. Noncoated membrane invaginations are involved in binding and internalization of cholera and tetanus toxins. Nature 296:651-653 (1982) https://doi.org/10.1038/296651a0
  12. Schnitzer, J. E., Liu, J. & Oh, P. Endothelial caveolae have the molecular transport machinery for vesicle budding, docking, and fusion including VAMP, NSF, SNAP, annexins, and GTPases. J Biol Chem 270: 14399-14404 (1995) https://doi.org/10.1074/jbc.270.24.14399
  13. Rothberg, K. G., Ying, Y. S., Kolhouse, J. F., Kamen, B. A. & Anderson, R. G. The glycophospholipidlinked folate receptor internalizes folate without entering the clathrin-coated pit endocytic pathway. J Cell Biol 110:637-649 (1990) https://doi.org/10.1083/jcb.110.3.637
  14. Anderson, R. G., Kamen, B. A., Rothberg, K. G. & Lacey, S. W. Potocytosis: sequestration and transport of small molecules by caveolae. Science 255:410-411 (1992) https://doi.org/10.1126/science.1310359
  15. Engelman, J. A. et al. Molecular genetics of the caveolin gene family: implications for human cancers, diabetes, Alzheimer disease, and muscular dystrophy. Am J Hum Genet 63:1578-1587 (1998) https://doi.org/10.1086/302172
  16. Smart, E. J. et al. Caveolins, liquid-ordered domains, and signal transduction. Mol Cell Biol 19:7289-7304 (1999) https://doi.org/10.1128/MCB.19.11.7289
  17. Nystrom, F. H., Chen, H., Cong, L. N., Li, Y. & Quon, M. J. Caveolin-1 interacts with the insulin receptor and can differentially modulate insulin signaling in transfected Cos-7 cells and rat adipose cells. Mol Endocrinol 13:2013-2024 (1999) https://doi.org/10.1210/me.13.12.2013
  18. Williams, T. M. & Lisanti, M. P. Caveolin-1 in oncogenic transformation, cancer, and metastasis. Am J Physiol Cell Physiol 288:C494-506 (2005) https://doi.org/10.1152/ajpcell.00458.2004
  19. Priuska, E. M. & Schacht, J. Formation of free radicals by gentamicin and iron and evidence for an iron/ gentamicin complex. Biochem Pharmacol 50:1749-1752 (1995) https://doi.org/10.1016/0006-2952(95)02160-4
  20. Segal, J. A. & Skolnick, P. Polyamine-like actions of aminoglycosides and aminoglycoside derivatives at NMDA receptors. Eur J Pharmacol 347:311-317 (1998) https://doi.org/10.1016/S0014-2999(98)00108-3
  21. Nagy, I., Bodmer, M., Brors, D. & Bodmer, D. Early gene expression in the organ of Corti exposed to gentamicin. Hear Res 195:1-8 (2004) https://doi.org/10.1016/j.heares.2004.04.010
  22. Dehne, N., Rauen, U., de Groot, H. & Lautermann, J. Involvement of the mitochondrial permeability transition in gentamicin ototoxicity. Hear Res 169:47-55 (2002) https://doi.org/10.1016/S0378-5955(02)00338-6
  23. Lawlor, P., Marcotti, W., Rivolta, M. N., Kros, C. J. & Holley, M. C. Differentiation of mammalian vestibular hair cells from conditionally immortal, postnatal supporting cells. J Neurosci 19(21):9445-9458 (1999) https://doi.org/10.1523/JNEUROSCI.19-21-09445.1999
  24. Hackett, L. et al. E-cadherin and the differentiation of mammalian vestibular hair cells. Exp Cell Res 278: 19-30 (2002) https://doi.org/10.1006/excr.2002.5574
  25. Rivolta, M. N. & Holley, M. C. Cell lines in inner ear research. J Neurobiol 53(2):306-318 (2002) https://doi.org/10.1002/neu.10111
  26. Cohen, A. W., Hnasko, R., Schubert, W. & Lisanti, M. P. Role of caveolae and caveolins in health and disease. Physiol Rev 84:1341-1379 (2004) https://doi.org/10.1152/physrev.00046.2003
  27. Williams, T. M. & Lisanti, M. P. The Caveolin genes: from cell biology to medicine. Ann Med 36:584-595 (2004) https://doi.org/10.1080/07853890410018899
  28. Takumida, M. & Anniko, M. Nitric oxide in the inner ear. Curr Opin Neurol 15:11-15 (2002) https://doi.org/10.1097/00019052-200202000-00003
  29. Takumida, M. & Anniko, M. Detection of nitric oxide in the guinea pig inner ear, using a combination of aldehyde fixative and 4,5-diaminofluorescein diacetate. Acta Otolaryngol 121:460-464 (2001) https://doi.org/10.1080/00016480121093
  30. Takumida, M. & Anniko, M. Nitric oxide in guinea pig vestibular sensory cells following gentamicin exposure in vitro. Acta Otolaryngol 121:346-350 (2001) https://doi.org/10.1080/000164801300102734
  31. Tuper, G., Ahmad, N. & Seidman, M. Mechanisms of Ototoxicity. Perspectives on Hearing and Hearing Disorder. Research and Diagnostics 9: 2-10 (2005)