Skip to main content
PMC home page
JARO: Journal of the Association for Research in Otolaryngology logoLink to JARO: Journal of the Association for Research in Otolaryngology
. 2003 Jan 15;4(2):176–195. doi: 10.1007/s10162-002-2036-8

Distribution of Gentamicin in the Guinea Pig Inner Ear after Local or Systemic Application

Shun-ichi Imamura 1, Joe C Adams 2,✉
PMCID: PMC3202710  PMID: 12943372

Abstract

Uptake and retention of gentamicin by cells in the guinea pig inner ear after a single peritoneal injection or local application on the round window were investigated using immunocytochemistry to localize the drug. The cells that accumulated the drug under the two conditions were the same, but staining for the drug was more intense and was often accompanied by widespread cochlear degeneration following local application. Soon after drug administration by either route, there was diffuse staining for the drug throughout all tissue within the labyrinth, including bone. At later times when distinct cell staining became evident, virtually all cell types were found to be positive, with several cell types staining more darkly for the drug than hair cells, indicating that hair cells were not the most avid in accumulating gentamicin. The infracuticular portion of auditory and vestibular hair cells as well as type III fibrocytes of the spiral ligament were positively stained in almost all cases and these sites were found to be positive for as long as six months post administration. In animals with loss of the organ of Corti, there was unusually intense staining for gentamicin in root cells of the spiral ligament, in marginal cells of the stria vascularis, and in cells of the spiral limbus. Dark staining of surviving cells in cases with overt tissue destruction suggests that variability in the extent of damage caused by the drug was determined more by the degree of its local uptake than by differences in animals' capacities to metabolize the drug systemically. The present results show that gentamicin may damage or destroy all cochlear cells following a single round window application. The findings broaden the scope of our knowledge of cochlear gentamicin uptake and damage and have implications for treatment of patients with vestibular disorders by infusion of aminoglycosides into the middle ear, as well as implications for prospects of rehabilitating patients that have been deafened by aminoglycosides.

Keywords: aminoglycoside, immunocytochemistry, inner ear, ototoxicity

Full Text

The Full Text of this article is available as a PDF (809.5 KB).

Acknowledgements

This work was supported by NIH grant DC 03929. The authors are grateful to M.C. Liberman, J.B. Nadol, Jr., and W.F. Sewell for helpful comments on the manuscript.

References

  • 1.Adams JC. Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains. J. Histochem. Cytochem. 1992;40:1457–1463. doi: 10.1177/40.10.1527370. [DOI] [PubMed] [Google Scholar]
  • 2.Aran JM, Erre JP, Lima da Costa D, Debbarh I, Dulon D. Acute and chronic effects of aminoglycosides on cochlear hair cells. Ann. N. Y. Acad. Sci. 1999;884:60–68. doi: 10.1111/j.1749-6632.1999.tb08636.x. [DOI] [PubMed] [Google Scholar]
  • 3.Balogh K, Hiraide F, Ishi D. Distribution of radioactive dihydrostreptomycin in the cochlea—an autoradiographic study. Ann. Otol. (St. Louis) 1970;79:641–652. doi: 10.1177/000348947007900329. [DOI] [PubMed] [Google Scholar]
  • 4.Blakley BW. Update on intratympanic gentamicin for Meniere's disease. Laryngoscope. 2000;110:236–240. doi: 10.1097/00005537-200002010-00009. [DOI] [PubMed] [Google Scholar]
  • 5.Brummett RE, Traynor J, Brown R, Himes D. Cochlear damage resulting from kanamycin and furosemide. Acta Otolaryngol. 1975;80:86–92. doi: 10.3109/00016487509121305. [DOI] [PubMed] [Google Scholar]
  • 6.Bryant GM, Cronin–Schreiber R, Alexander A, Norris CH, Quine DB, Guth PS. The potentiation of ototoxicity when aminooxyacetic acid and kanamycin are co-administered. Hear. Res. 1984;15:173–178. doi: 10.1016/0378-5955(84)90048-0. [DOI] [PubMed] [Google Scholar]
  • 7.Conlon BJ, McSwain SD, Smith DW. Topical gentamicin and ethacrynic acid: effects on cochlear function. Laryngoscope. 1998;108:1087–1089. doi: 10.1097/00005537-199807000-00024. [DOI] [PubMed] [Google Scholar]
  • 8.Cotanche DA. Structural recovery from sound and aminoglycoside damage in the avian cochlea. Audiol. Neurootol. 1999;4:271–285. doi: 10.1159/000013852. [DOI] [PubMed] [Google Scholar]
  • 9.DeCicco MJ, Hoffer ME, Kopke RD, Wester D, Allen KA, Gottshall K, O'Leary MJ. Round-window microcatheter-administered microdose gentamicin: results from treatment of tinnitus associated with Meniere's disease. Int. Tinnitus J. 1998;4:141–143. [PubMed] [Google Scholar]
  • 10.DeGroot JCMJ, Meeuwsen F, Ruizentaal WE, Veldman JE. Ultrastructural localization of gentamicin. Hear. Res. 1990;50:35–42. doi: 10.1016/0378-5955(90)90031-J. [DOI] [PubMed] [Google Scholar]
  • 11.Dulon D, Aran JM, Zajic G, Schacht J. Comparative uptake of gentamicin, netilmicin, and amikacin in the guinea pig cochlea and vestibule. Antimicrob. Agents Chemother. 1986;30:96–100. doi: 10.1128/aac.30.1.96. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Dulon D, Hiel H, Aurousseau C, Erre JP, Aran JM. Pharmacokinetics of gentamicin in the sensory hair cells of the organ of Corti: rapid uptake and long term persistence. C. R. Acad. Sci. III. 1993;316:682–687. [PubMed] [Google Scholar]
  • 13.Federspil P. Pharmacokinetics of aminoglycoside antibiotics in the perilymph. In: Lerner SA, Mats GJ, Hawkins Jr. JE, editors. Aminoglycoside Ototoxicity. Little Brown: Boston; 1981. pp. 99–108. [Google Scholar]
  • 14.Forge A, Schacht J. Aminoglycoside antibiotics. Audiol. Neurootol. 2000;5:3–22. doi: 10.1159/000013861. [DOI] [PubMed] [Google Scholar]
  • 15.Gao WQ. Therapeutic potential of neurotrophins for treatment of hearing loss. Mol. Neurobiol. 1998;17:17–31. doi: 10.1007/BF02802022. [DOI] [PubMed] [Google Scholar]
  • 16.Hashino E, Shero M. Endocytosis of aminoglycoside antibiotics in sensory hair cells. Brain Res. 1995;704:135–140. doi: 10.1016/0006-8993(95)01198-6. [DOI] [PubMed] [Google Scholar]
  • 17.Hashino E, Shero M, Salvi RJ. Lysosomal targeting and accumulation of aminoglycoside antibiotics in sensory hair cells. Brain Res. 1997;777:75–85. doi: 10.1016/S0006-8993(97)00977-3. [DOI] [PubMed] [Google Scholar]
  • 18.Hawkins Jr JE. Comparative otopathology: aging, noise, and ototoxic drugs. Adv. Otorhinolaryngol. 1973;20:125–141. doi: 10.1159/000393093. [DOI] [PubMed] [Google Scholar]
  • 19.Hawkins Jr JE, Johnsson LG. Histopathology of cochlear and vestibular ototoxicity. In: Lerner SA, Mats GJ, Hawkins Jr JE, editors. Aminoglycoside ototoxicity. Boston: Little Brown; 1981. pp. 175–195. [Google Scholar]
  • 20.Hayashida T, Nomura Y, Iwamori M, Nagai Y, Kurata T. Distribution of gentamicin by immunofluorescence in the guinea pig inner ear. Arch. Otorhinolaryngol. 1985;242:257–264. doi: 10.1007/BF00453548. [DOI] [PubMed] [Google Scholar]
  • 21.Hayashida T. An immunohistochemical method for the study of aminoglycoside ototoxicity in the guinea pig cochlea using decalcified frozen sections. Arch. Otorhinolaryngol. 1989;246:161–164. doi: 10.1007/BF00456659. [DOI] [PubMed] [Google Scholar]
  • 22.Hayashida T, Hiel H, Dulon D, Erre JP, Guihaume A, Aran JM. Dynamic changes following combined treatment with gentamicin and ethacrynic acid with and without acoustic stimulation. Acta Otolaryngol. (Stockh.) 1989;108:404–413. doi: 10.3109/00016488909125546. [DOI] [PubMed] [Google Scholar]
  • 23.Hiel H, Bennani H, Erre J, Aurousseau C, Aran J. Kinetics of gentamicin in cochlear hair cells after chronic treatment. Acta Otolaryngol. (Stockh.) 1992a;112:272–277. doi: 10.1080/00016489.1992.11665417. [DOI] [PubMed] [Google Scholar]
  • 24.Hiel H, Schamel A, Erre J, Hayashida T, Dulon D, Aran J. Cellular and subcellular localization of tritiated gentamicin in the guinea pig cochlea following combined treatment with ethacrynic acid. Hear. Res. 1992b;57:157–165. doi: 10.1016/0378-5955(92)90148-g. [DOI] [PubMed] [Google Scholar]
  • 25.Hiel H, Erre J, Aurousseau C, Bouali R, Dulon D, Aran J. Gentamicin uptake by cochlear hair cells precedes hearing impairment during chronic treatment. Audiology. 1993;32:78–87. doi: 10.3109/00206099309072930. [DOI] [PubMed] [Google Scholar]
  • 26.Husmann KR, Morgan AS, Girod DA, Durham D. Round window administration of gentamicin: a new method for the study of ototoxicity of cochlear hair cells. Hear. Res. 1998;125:109–119. doi: 10.1016/S0378-5955(98)00137-3. [DOI] [PubMed] [Google Scholar]
  • 27.Ichimiya I, Adams JC, Kimura R. Changes in immunostaining of cochleas with experimentally induced endolymphatic hydrops. Ann. Otol. Rhinol. Laryngol. 1994;103:457–468. doi: 10.1177/000348949410300607. [DOI] [PubMed] [Google Scholar]
  • 28.Imamura S, Adams JC. Immunolocalization of peptide 19 and other calcium-binding proteins in the guinea pig cochlea. Anat. Embryol. (Berl.) 1996;194:407–418. doi: 10.1007/BF00198543. [DOI] [PubMed] [Google Scholar]
  • 29.Iwamori M, Tayama N, Nomura Y, Nagai Y. Hormone-dependent enhancement in binding of oto- and nephrotoxic aminoglycoside antibiotics. Acta Otolaryngol. Suppl. (Stockh.) 1994;514:117–121. doi: 10.3109/00016489409127574. [DOI] [PubMed] [Google Scholar]
  • 30.Kikuchi T, Kimura RS, Adams JC. Gap junctions in the rat cochlea: immunohistochemical and ultrastructural analysis. Anat. Embryol. (Berl.) 1995;191:101–118. doi: 10.1007/BF00186783. [DOI] [PubMed] [Google Scholar]
  • 31.Kimura RS, Lee K-S, Nye CL, Trehey JA. Effects of systemic and lateral semicircular canal administration of aminoglycosides on normal and hydropic inner ears. Acta Otolaryngol. (Stockh.) 1991;111:1021–1030. doi: 10.3109/00016489109100751. [DOI] [PubMed] [Google Scholar]
  • 32.Leake PA, Snyder RL. Uptake of horseradish peroxidase from perilymph by cochlear hair cells. Hear. Res. 1987;25:153–171. doi: 10.1016/0378-5955(87)90088-8. [DOI] [PubMed] [Google Scholar]
  • 33.Marzella PL, Clark GM. Growth factors, auditory neurones and cochlear implants: a review. Acta Otolaryngol. 1999;119:407–412. doi: 10.1080/00016489950180919. [DOI] [PubMed] [Google Scholar]
  • 34.Meyer J, Mack AL, Gummer AW. Pronounced infracuticular endocytosis in mammalian outer hair cells. Hear. Res. 2001;161:10–22. doi: 10.1016/S0378-5955(01)00338-0. [DOI] [PubMed] [Google Scholar]
  • 35.Mugnaini E, Dahl AL. Zinc-aldehyde fixation for light-microscopic immunocytochemistry of nervous tissues. J. Histochem. Cytochem. 1983;31:1435–1438. doi: 10.1177/31.12.6355290. [DOI] [PubMed] [Google Scholar]
  • 36.Nadol JB, Adams JC, Kim JR. Degenerative changes in the organ of Corti and lateral cochlear wall in experimental endolymphatic hydrops and human Meniere's disease. Acta Otolaryngol. Suppl. (Stockh.) 1995;519:47–59. doi: 10.3109/00016489509121870. [DOI] [PubMed] [Google Scholar]
  • 37.Nakashima T, Teranishi M, Hibi T, Kobayashi M, Umemura M. Vestibular and cochlear toxicity of aminoglycosides—a review. Acta Otolaryngol. 2000;120:904–911. doi: 10.1080/00016480050218627. [DOI] [PubMed] [Google Scholar]
  • 38.Portmann M, Darrouzet J, Coste C. Distribution within the cochlea of dihydrostreptomycin injected into the circulation. Arch. Otolaryngol. (Chicago). 1974;100:473–475. doi: 10.1001/archotol.1974.00780040487014. [DOI] [PubMed] [Google Scholar]
  • 39.Richardson GP, Forge A, Cros CJ, Fleming J, Brown SD, Steel KP. Myosin VIIA is required for aminoglycoside accumulation in cochlear hair cells. J. Neurosci. 1997;17:9506–9519. doi: 10.1523/JNEUROSCI.17-24-09506.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Ryback LP. Ototoxic mechanisms. In: Altschuler RA, Bobbin RP, Hoffman DW, editors. Neurobiology of hearing: the cochlea. New York: Raven Press; 1986. pp. 442–454. [Google Scholar]
  • 41.Schacht J. Molecular mechanisms of drug-induced hearing loss. Hear. Res. 1986;22:297–304. doi: 10.1016/0378-5955(86)90105-X. [DOI] [PubMed] [Google Scholar]
  • 42.Schacht J. Biochemical basis of aminoglycoside ototoxicity. Otolaryngol. Clin. N. Am. 1993;26:845–856. [PubMed] [Google Scholar]
  • 43.Schulte BA, Adams JC. Distribution of immunoreactive Na+, K+-ATPase in gerbil cochlea. J. Histochem Cytochem. 1989;37:127–134. doi: 10.1177/37.2.2536055. [DOI] [PubMed] [Google Scholar]
  • 44.Siegel JH, Brownell WE. Synaptic and golgi membrane recycling in cochlear hair cells. J. Neurocytol. 1986;15:311–328. doi: 10.1007/BF01611434. [DOI] [PubMed] [Google Scholar]
  • 45.Spicer SS, Schulte BA. Golgi-canalicular reticulum system in ion transporting fibrocytes and outer sulcus epithelium of gerbil cochlea. Anat. Rec. 1997;249:117–127. doi: 10.1002/(SICI)1097-0185(199709)249:1<117::AID-AR14>3.3.CO;2-W. [DOI] [PubMed] [Google Scholar]
  • 46.Spicer SS, Thomopoulos GN, Schulte BA. Cytologic evidence for mechanisms of K+ transport and genesis of Hensen bodies and subsurface cisternae in outer hair cells. Anat. Rec. 1998;251:97–113. doi: 10.1002/(SICI)1097-0185(199805)251:1<97::AID-AR15>3.0.CO;2-6. [DOI] [PubMed] [Google Scholar]
  • 47.Stone JS, Rubel EW. Cellular studies of auditory hair cell regeneration in birds. Proc. Natl. Acad. Sci. U S A. 2000;97:11714–11721. doi: 10.1073/pnas.97.22.11714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 48.Tachibana M, Morioka H, Machino M, Mizukoshi O. Binding sites of an aminoglycoside in the cochlea examined by immunocytochemistry. Histochemistry. 1985;83:237–240. doi: 10.1007/BF00953990. [DOI] [PubMed] [Google Scholar]
  • 49.Takada A, Bledsoe Jr S, Schacht J. An energy-dependent step in aminoglycoside ototoxicity: prevention of gentamicin ototoxicity during reduced endolymphatic potential. Hear. Res. 1985;19:245–251. doi: 10.1016/0378-5955(85)90144-3. [DOI] [PubMed] [Google Scholar]
  • 50.Tran Ba Huy P, Manuel C, Meulemans A. Kinetics of aminoglycoside antibiotics in perilymph and endolymph in animals. In: Lerner SA, Mats GJ, Hawkins Jr JE, editors. Aminoglycoside ototoxicity. Boston: Little, Brown; 1981a. pp. 81–97. [Google Scholar]
  • 51.Tran Ba Huy P, Manuel C, Meulemans A, Sterkers O, Amiel C. Pharmacokinetics of gentamicin in perilymph and endolymph of the rat as determined by radioimmunoassay. J. Infect. Dis. 1981b;143:476–486. doi: 10.1093/infdis/143.3.476. [DOI] [PubMed] [Google Scholar]
  • 52.Tran Ba Huy P, Meulemans A, Wassef M, Manuel C, Sterkers O, Amiel C. Gentamicin persistent in rat endolymph and perilymph after two-day constant infusion. Antimicrob. Agents Chemther. 1983;23:344–346. doi: 10.1128/aac.23.2.344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 53.Tran Ba Huy P, Bernard P, Schacht J. Kinetics of gentamicin uptake and release in the rat; comparison of inner ear tissues and fluids with others organs. J. Clin. Invest. 1986;77:1492–1500. doi: 10.1172/JCI112463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 54.Veldman JE, Meeuwsen F, van Dijk M, Key Q, Huizing EH. Progress in temporal bone histopathology. II. Immuno-technology applied to the temporal bone. Acta Otolaryngol. Suppl. 1985;423:29–35. doi: 10.3109/00016488509122909. [DOI] [PubMed] [Google Scholar]
  • 55.von Ilberg C, Spoendlin H, Arnold W. Autoradiographical distribution of locally applied dihydrostreptomycin in the inner ear. Acta Otolaryngol. (Stockh.) 1971;71:159–165. doi: 10.3109/00016487109125345. [DOI] [PubMed] [Google Scholar]
  • 56.Wersall J, Lundquist PG, Bjorkroth B. Ototoxicity of gentamicin. J. Infect. Dis. 1969;119:410–416. doi: 10.1093/infdis/119.4-5.410. [DOI] [PubMed] [Google Scholar]
  • 57.Wersall J. Ototoxic antibiotics: a review. Acta Otolaryngol. Suppl. 1995;519:26–29. doi: 10.3109/00016489509121866. [DOI] [PubMed] [Google Scholar]
  • 58.Williams SE, Zenner HP, Schacht J. Three molecular steps aminoglycoside ototoxicity demonstrated in outer hair cells. Hear. Res. 1987;30:11–18. doi: 10.1016/0378-5955(87)90177-8. [DOI] [PubMed] [Google Scholar]
  • 59.Yamane H, Nakai Y, Konishi K. Furosemide-induced alteration of drug pathway to cochlea. Acta Otolaryngol. Suppl. 1988;447:28–35. doi: 10.3109/00016488809102854. [DOI] [PubMed] [Google Scholar]

Articles from JARO: Journal of the Association for Research in Otolaryngology are provided here courtesy of Association for Research in Otolaryngology

RESOURCES