Clin Exp Otorhinolaryngol.  2018 Jun;11(2):109-117. 10.21053/ceo.2017.00857.

Impact of Nicotine Exposure on Hair Cell Toxicity and Embryotoxicity During Zebrafish Development

Affiliations
  • 1Department of Otorhinolaryngology-Head and Neck Surgery, Kyungpook National University School of Medicine, Daegu, Korea.
  • 2Department of Otorhinolaryngology-Head and Neck Surgery, Korea University College of Medicine, Seoul, Korea. mednlaw@korea.ac.kr
  • 3Biomedical Research Center, Korea University Ansan Hospital, Ansan, Korea.

Abstract


OBJECTIVES
Nicotine has various adverse effects including negative impacts associated with maternal exposure. In the current study, we examined nicotine-induced damage of hair cells and embryotoxicity during zebrafish development.
METHODS
Zebrafish embryos were exposed to nicotine at several concentrations (5, 10, 20, and 40 μM) and embryotoxicity were evaluated at 72 hours, including hatching rate, mortality, teratogenicity rate, and heart rate. Hair cells within the supraorbital (SO1 and SO2), otic (O1), and occipital (OC1) neuromasts were identified at 120 hours. Apoptosis and mitochondrial damage of hair cells were analyzed using TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling) and DASPEI (2-[4-(dimethylamino)styryl]-N-ethylpyridinium iodide) assays, respectively, and changes of ultrastructure were observed by scanning electron microscopy.
RESULTS
The control group without nicotine appeared normal with overall mortality and teratogenicity rate < 5%. The hatching rate and mortality rate was not significantly different according to nicotine concentration (n=400 each). The abnormal morphology rate (n=400) increased and heart rate (n=150) decreased with increasing nicotine concentration (P < 0.05). Nicotine-induced hair cell damage significantly increased as nicotine concentration increased. A significantly greater number of TUNEL-positive cells (P < 0.01) and markedly smaller DASPEI area (P < 0.01) were shown as nicotine concentration increased.
CONCLUSION
The current results suggest that nicotine induces dose-dependent hair cell toxicity in embryos by promoting apoptosis and mitochondrial and structural damage.

Keyword

Nicotine; Tobacco; Zebrafish; Hair Cells; Ototoxicity; Embryotoxicity

MeSH Terms

Apoptosis
Embryonic Structures
Female
Hair*
Heart Rate
In Situ Nick-End Labeling
Maternal Exposure
Microscopy, Electron, Scanning
Mortality
Nicotine*
Tobacco
Zebrafish*
Nicotine
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