Uptake of Gentamicin by Bullfrog Saccular Hair Cells in vitro

Peter Steyger, S. L. Peters, J. Rehling, A. Hordichok, C. F. Dai

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Vertebrate sensory hair cells in the inner ear are pharmacologically sensitive to aminoglycoside antibiotics. Although the ototoxicity, of aminoglycosides is well known, the route of drug uptake by hair cells and mechanisms of cytotoxicity remain poorly understood. Previously published studies have documented the intracellular distribution of gentamicin using immunocytochemical, electron microscopic, and autoradiographic methods. In this article, we compare the subcellular distribution of fluorescently conjugated gentamicin (gentamicin-Texas Red, GTTR) with immunolabeled gentamicin using confocal or electron microscopy. Gentamicin (detected by post-fixation immunocytochemistry) and GTTR were rapidly taken up by hair cells throughout the bullfrog saccular explant in vitro and preferentially in peripheral hair cells. Immunolabeled gentamicin and GTTR were observed at the apical membranes of hair cells, particularly in their hair bundles. GTTR was also identified within a variety of subcellular compartments within hair cells, including lysosomes, mitochondria, Golgi bodies, endoplasmic reticulum, and nuclei, and in similar structures by immunoelectron microscopy. The distributions of GTTR and immunolabeled gentamicin are largely identical and corroborate a variety of published immunocytochemical and autoradiography studies. Thus, GTTR is a valid fluorescent probe with which to investigate the pharmacokinetics and mechanisms of gentamicin accumulation.

Original languageEnglish (US)
Pages (from-to)565-578
Number of pages14
JournalJARO - Journal of the Association for Research in Otolaryngology
Volume4
Issue number4
DOIs
StatePublished - Dec 2003

Fingerprint

Rana catesbeiana
Gentamicins
Aminoglycosides
In Vitro Techniques
Immunoelectron Microscopy
Inner Ear
Lysosomes
Autoradiography
Fluorescent Dyes
Confocal Microscopy
Endoplasmic Reticulum
Vertebrates
Texas red

Keywords

  • Drug accumulation
  • Endocytosis
  • Gentamicin
  • Hair cells
  • Ototoxicity

ASJC Scopus subject areas

  • Otorhinolaryngology
  • Physiology

Cite this

Uptake of Gentamicin by Bullfrog Saccular Hair Cells in vitro. / Steyger, Peter; Peters, S. L.; Rehling, J.; Hordichok, A.; Dai, C. F.

In: JARO - Journal of the Association for Research in Otolaryngology, Vol. 4, No. 4, 12.2003, p. 565-578.

Research output: Contribution to journalArticle

Steyger, Peter ; Peters, S. L. ; Rehling, J. ; Hordichok, A. ; Dai, C. F. / Uptake of Gentamicin by Bullfrog Saccular Hair Cells in vitro. In: JARO - Journal of the Association for Research in Otolaryngology. 2003 ; Vol. 4, No. 4. pp. 565-578.
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