microRNA-183 is Essential for Hair Cell Regeneration after Neomycin Injury in Zebrafish
10.3349/ymj.2018.59.1.141
- Author:
Chang Woo KIM
1
;
Ji Hyuk HAN
;
Ling WU
;
Jae Young CHOI
Author Information
1. Department of Otorhinolaryngology, Hallym University College of Medicine, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
microRNA;
ototoxicity;
regeneration;
zebrafish
- MeSH:
Animals;
Animals, Genetically Modified;
Cell Count;
Gene Expression Profiling;
Gene Expression Regulation/drug effects;
Gene Knockdown Techniques;
Green Fluorescent Proteins/metabolism;
Hair Cells, Auditory/drug effects;
Hair Cells, Auditory/physiology;
Larva/drug effects;
Larva/genetics;
MicroRNAs/genetics;
MicroRNAs/metabolism;
Morpholinos/pharmacology;
Neomycin/toxicity;
Regeneration/drug effects;
Regeneration/genetics;
Zebrafish/genetics
- From:Yonsei Medical Journal
2018;59(1):141-147
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: microRNAs (miRNAs) are non-coding RNAs composed of 20 to 22 nucleotides that regulate development and differentiation in various organs by silencing specific RNAs and regulating gene expression. In the present study, we show that the microRNA (miR)-183 cluster is upregulated during hair cell regeneration and that its inhibition reduces hair cell regeneration following neomycin-induced ototoxicity in zebrafish. MATERIALS AND METHODS: miRNA expression patterns after neomycin exposure were analyzed using microarray chips. Quantitative polymerase chain reaction was performed to validate miR-183 cluster expression patterns following neomycin exposure (500 µM for 2 h). After injection of an antisense morpholino (MO) to miR-183 (MO-183) immediately after fertilization, hair cell regeneration after neomycin exposure in neuromast cells was evaluated by fluorescent staining (YO-PRO1). The MO-183 effect also was assessed in transgenic zebrafish larvae expressing green fluorescent protein (GFP) in inner ear hair cells. RESULTS: Microarray analysis clearly showed that the miR-183 cluster (miR-96, miR-182, and miR-183) was upregulated after neomycin treatment. We also confirmed upregulated expression of the miR-183 cluster during hair cell regeneration after neomycin-induced ototoxicity. miR-183 inhibition using MO-183 reduced hair cell regeneration in both wild-type and GFP transgenic zebrafish larvae. CONCLUSION: Our work demonstrates that the miR-183 cluster is essential for the regeneration of hair cells following ototoxic injury in zebrafish larvae. Therefore, regulation of the miR-183 cluster can be a novel target for stimulation of hair cell regeneration.
