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Study of the Mechanisms by Which Aminoglycoside Damage Is Prevented in Chick Embryonic Hair Cells

  • Huanju Bai
  • Xi Wang
  • Xue Gao
  • Jie Bing
  • Weiqian Wang
  • Xuebo Zhang
  • Chao Xi
  • Lingling Jiang
  • Xinwen Zhang
  • Zhongming Han
  • Shaoju ZengEmail author
  • Jincao XuEmail author
Research Article

Abstract

A major side effect of aminoglycoside antibiotics is mammalian hair cell death. It is thus intriguing that embryonic chick hair cells treated with aminoglycosides at embryonic day (E) 12 are insensitive to ototoxicity. To exclude some unknown factors in vivo that might be involved in preventing aminoglycoside damage to embryonic hair cells, we first cultured chick embryonic basilar papilla (BP) with an aminoglycoside antibiotic in vitro. The results indicated that the hair cells were almost intact at E12 and E14 and were only moderately damaged in most parts of the BP at E16 and E18. Generally, hair cells residing in the approximate and abneural regions were more susceptible to streptomycin damage. After incubation with gentamicin-conjugated Texas Red (GTTR), which is typically used to trace the entry route of aminoglycosides, GTTR fluorescence was not remarkable in hair cells at E12, was weak at E14, but was relatively strong in the proximal part of BP at E18. This result indicates that the amounts of GTTR that entered the hair cells are related to the degrees of aminoglycoside damage. The study further showed that the fluorescence intensity of GTTR decreased to a low level at E14 to E18 after disruption of mechanotransduction machinery, suggesting that the aminoglycoside entry into hair cells was mainly through mechanotransduction channels. In addition, most of the entered GTTR was not found to be colocalized with mitochondria even at E18. This finding provides another reason to explain why embryonic chick hair cells are insensitive to aminoglycoside damage.

Keywords

antibiotic toxicity mechanotransduction channels transient receptor potential ion channel chicken cochlea mitochondria 

Abbreviations

BP

Basilar papilla

GTTR

Gentamicin-conjugated Texas Red

MET

Mechano-electrical transduction

TRPA1

Transient receptor potential (TRP) ion channel 1

BAPTA

2-bis (o-aminophenoxy) ethane-N, N, N′, N′-tetra-acetic acid

FM1–43

(N- (3-triethylammoniumpropyl)- 4- (4-(dibutylamino) styryl) pyridiniumdibromide

Notes

Acknowledgments

We thank Dr. Jin-Liu in the Experimental Technology Center for Life Sciences and the College of Life Sciences, Beijing Normal University, for technological assistance.

Funding Information

This work was supported by the National Natural Science Foundation of China through grants to ZM Han (No. 81371352), SJ Zeng (Nos. 31672283 and 31372200), XW Zhang (No. 31560275 and 31360517), and XB Zhang (No. 31360243).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no competing interests.

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Copyright information

© Association for Research in Otolaryngology 2018

Authors and Affiliations

  1. 1.Beijing Key Laboratory of Gene Resource and Molecular DevelopmentBeijing Normal UniversityBeijingChina
  2. 2.Department of OtorhinolaryngologyThe General Hospital of the PLA Rocket ForceBeijingChina
  3. 3.Ministry of Education Key Laboratory for Ecology of Tropical Islands, College of Life SciencesHainan Normal UniversityHaikouChina

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