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microRNA-183 is involved in the differentiation and regeneration of Notch signaling-prohibited hair cells from mouse cochlea

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      Auditory hair cell regeneration following injury is critical to hearing restoration. The Notch signaling pathway participates in the regulation of inner ear development and cell differentiation. Recent evidence suggests that microRNA (miR)-183 has a similar role in the inner ear. However, it is unclear how Notch signaling functions in hair cell regeneration in mammals and if there is cross-talk between Notch signaling and miR-183. The present study used a gentamicin-induced cochlear injury mouse model. Gentamicin-induced damage of the hair cells activated the Notch signaling pathway and downregulated miR-183 expression. Notch signaling inhibition by the γ-secretase inhibitor, 24-diamino-5-phenylthiazole (DAPT), attenuated gentamicin-induced hair cell loss and reversed the downregulation of miR-183 expression. Further investigation revealed that the novel hair cells produced, induced by DAPT, were derived from transdifferentiated supporting cells. Additionally, myosin VI-positive hair cell numbers were increased by Notch signaling inhibition in in vitro experiments with cultured neonatal mouse inner ear precursor cells. This effect was reversed by miR-183 inhibition. These findings indicate that the Notch signaling pathway served a repressing role during the regeneration of hair cells. Inhibiting this signal improved hair cell regeneration in the gentamicin-damaged cochlear model. miR-183 was demonstrated to be involved in hair cell differentiation and regeneration, and was required for the differentiation of the Notch-inhibited hair cells.

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      Most cited references 21

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        A novel microRNA (miRNA) quantification method has been developed using stem–loop RT followed by TaqMan PCR analysis. Stem–loop RT primers are better than conventional ones in terms of RT efficiency and specificity. TaqMan miRNA assays are specific for mature miRNAs and discriminate among related miRNAs that differ by as little as one nucleotide. Furthermore, they are not affected by genomic DNA contamination. Precise quantification is achieved routinely with as little as 25 pg of total RNA for most miRNAs. In fact, the high sensitivity, specificity and precision of this method allows for direct analysis of a single cell without nucleic acid purification. Like standard TaqMan gene expression assays, TaqMan miRNA assays exhibit a dynamic range of seven orders of magnitude. Quantification of five miRNAs in seven mouse tissues showed variation from less than 10 to more than 30 000 copies per cell. This method enables fast, accurate and sensitive miRNA expression profiling and can identify and monitor potential biomarkers specific to tissues or diseases. Stem–loop RT–PCR can be used for the quantification of other small RNA molecules such as short interfering RNAs (siRNAs). Furthermore, the concept of stem–loop RT primer design could be applied in small RNA cloning and multiplex assays for better specificity and efficiency.
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          In situ detection of miRNAs in animal embryos using LNA-modified oligonucleotide probes.

          MicroRNAs (miRNAs) are 20-23 nucleotide (nt) RNA molecules that regulate gene expression post-transcriptionally. A key step toward understanding the function of the hundreds of miRNAs identified in animals is to determine their expression during development. Here we performed a detailed analysis of conditions for in situ detection of miRNAs in the zebrafish embryo using locked nucleic acid (LNA)-modified DNA probes and report expression patterns for 15 miRNAs in the mouse embryo.

            Author and article information

            [1 ]Department of Otolaryngology, The First Affiliated Hospital and Institute of Otorhinolaryngology, Sun Yat-sen University, Guangzhou, Guangdong 510080, P.R. China
            [2 ]Department of Otolaryngology, People's Hospital of Meishan, Meishan, Sichuan 620010, P.R. China
            [3 ]Department of Otolaryngology, West China Hospital of Sichuan University, Chengdu, Sichuan 6100041, P.R. China
            [4 ]Department of Otolaryngology, People's Hospital of Dongguan, Dongguan, Guangdong 510080, P.R. China
            [5 ]Department of Otorhinolaryngology, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei 050000, P.R. China
            [6 ]Department of Otorhinolaryngology, The Second Affiliated Hospital, Guangzhou Medical University, Guangzhou, Guangdong 510260, P.R. China
            Author notes
            Correspondence to: Professor Hongyan Jiang, Department of Otolaryngology, The First Affiliated Hospital and Institute of Otorhinolaryngology, Sun Yat-sen University, 58 Zhongshan Road, Guangzhou, Guangdong 510080, P.R. China, E-mail: hongyanjiang@

            Contributed equally

            Mol Med Rep
            Mol Med Rep
            Molecular Medicine Reports
            D.A. Spandidos
            August 2018
            05 June 2018
            05 June 2018
            : 18
            : 2
            : 1253-1262
            Copyright: © Zhou et al.

            This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.



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