Expression and Regulatory Effect of miR-30b on Dynamin in Cochlear Hair Cells

LI Yang, TANG Xiao-in, YU Fei. et al

Abstract

To determine the expression of miR-30b in hair cells of mice, and its regulatory effect on the target gene DNM1 and expression of Dynamin, the key protein of synaptic endocytosis in inner hair cells. Methods The basilar membrane of cochlear in adult C57 mice was obtained. The expression of miR-30b in the hair cells was detected by in situ hybridization. Luciferase vector was constructed and transfected into 293T cells with miR-30b. Changes in luciferase activity were measured to verify whether DNM1 was the target gene of miR-30b. Adeno-associated virus carrying miR-30b were micro-injected into cochlear via the round window membrane. mRNA expressions of DNM1 and miR-30b were detected by RT-PCR 14 days later. The expression of Dynamin was detected by Western blot. Results miR-30b expressed in the inner and outer hair cells scattered in the region of the nucleus and cytoplasm. miR-30b reduced luciferase activity from the reporter vector containing DNM11 (PDNM1 and Dynamin were observed following transfection of AAV-miR-30b. Conclusion miR-30b expresses in inner and outer hair cells, which is consistent with the morphological orientation of dynamin. miR-30b inhibits the expression of Dynamin by targeting DNM11 gene.

 

Keywords: Hair cell miR-30b DNM1, Dynamin in situ hybridization, Dual luciferase reporter gene, Adeno-associated virus 

 

Full Text:

PDF


References


World Health Organization survey data for March 2018. [2018-02-11]. http://www. who. int/mediacentre/ factsheets/fs300/en/.

RAIMONDI A,FERGUSON SM.LOU X.et al. Overlapping role of dynamin isoforms in synaptic vesicle endocytosis. Neuron,2011,70(6):1100-1114.

ZARAGOSI LE. WDZIEKONSKI B. BRIGAND KL. et al. Small RNA sequencing reveals miR-642a-3p as a novel adipocyte-specific microRNA and miR-30 as a key regulator of human adipogenesis. Genome Biol.2011,12(7); R64[2018-02- 11]. https://doi.org/10. 1 l86/gb-20ll-12-7-r64.

WU T.ZHOU H, HONG Y,et al. MiR-30 family members negatively regulate osteoblast differentiation. J Biol Chem, 2012,287(10);7503-7511.

ZHONG X, LI N, LIANG S, et al. Identification of microRNAs regulating reprogramming factor LIN28 in embryonic stem cells and cancer cells. J Biol Chem,2010,285 (53);41961-41971.

MARTINEZ I, CAZALLA D. ALMSTEAD LL,et al. MiR- 29 and miR-30 regulate B-Myb expression during cellular senescence. Proc Natl Acad Sci U S A, 2011 , 108 ( 2 ); 522- 527.

LI J, DONATH S, LI Y, et al. MiR-30 regulates mitochondrial fission through targeting p53 and the dynamin- related protein-1 pathway. PLoS Genetics, 2010, 6 ( 1 ); el000795 [ 2018-02-11 ]. http;//journals. plos. org/plosgenetics/article/10. 1371 /journal, pgen. 1000795. doi: 10. 1371/journal, pgen. 1000795.

WANG XR, ZHANG XM. ZHEN J. et al. MicroRNA expression in the embryonic mouse inner ear. Neuroreport, 2010,21(9):611-617.

LILACH M, FRIEDMAN LM, KAREN B. MicroRNAs and epigenetic regulation in the mammalian inner ear;implications for deafness. Mamm Genome,2009,20(9/10):581-603.

SCHEFFER DI,SHEN J,COREY DP,et al. Gene expression by mouse inner ear hair cells during development. J Neurosci, 2015,35(16)s6366-6380.


Refbacks

  • There are currently no refbacks.