Autophagy-related Gene Expression of Organ of Corti in Aminoglycoside-treated Rats
Kim, Yeon Ju; Tian, Chunjie; Kim, Jangho; Kim, Young Sun; Shin, Beomyong; Lee, Jong Joo; Lim, Hye Jin; Kim, You-Sun; Choung, Yun-Hoon
Department of Otolaryngology, Ajou University School of Medicine
BACKGROUND: Autophagy is a major intracellular degradation process, by which cytoplasmic material is degraded via fusion of the double-membrane compartment, autophagosome with lysosomes. This process is important for the maintenance of cell homeostasis. Our previous data showed that enhanced autophagic flux can delay the aminoglycoside-induced ototoxicity, but the specific genes that are related with autophagy-dependent protective properties in hair cells have not been identified. To investigate the possible link between the autophagy process and hair cell protection against aminoglycoside-induced ototoxicity, we performed next generation sequencing (NGS) using ex vivo system.
METHODS: Organ of Corti explants of Sprague-Dawley rats (postnatal day 7) were cultured on tissue culture plates. The explants were exposed to: (a) Distilled water, (b) 50 μM gentamicin (GM) and (c) 50 μM GM + 50 pM rapamycin (RPM, as autophagy inducer) for 2 days. Transcriptome of organ of Corti cells were examined using RNA-sequencing. Differentially expressed genes were further verified for the organ of Corti with quantitative reverse transcription PCR (qRT-PCR).
RESULTS: First, we evaluated hair cell loss in the apical, middle and basal turns of the organ of Corti in each group. Immunohistochemical findings of organ of Cortis showed that the loss of stereocilia was much more in the GM-treated explants than in the GM + RPM-treated explants. Then the RNA samples were extracted from each group and RNA-sequencing was performed. We have shown the lists of differently expressed genes between samples. Results from the analysis are pending.
CONCLUSIONS: Autophagy may be closely connected with the aminoglycoside-mediated hair cell death and survival through the expression of specific genes.
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