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Expression of miR-140 in mouse inner ear hair cells and its regulation on synaptic protein Dynamin-1



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Expression of miR-140 in mouse inner ear hair cells and its regulation on synaptic protein Dynamin-1


TANG Xiaolin LI Yang YU Fei JIA Lifeng LI Huajun YUAN Wei

Department of Otolaryngology, Head and Neck Surgery, First Affiliated Hospital, Army Military Medical University (Third Military Medical University), Chongqing, 400038, China


miR-140 Dynamin-1 in situ hybridization inner hair cells adeno-associated virus

R322.923; R329.26; R394.3

Objective    To investigate the expression and distribution of miR-140 in mouse inner ear hair cells and its targeting regulation on Dnm1, the gene encoding synaptophysin Dynamin-1. Methods    After the cochlear basement membrane was harvested from healthy adult C57BL/6J mice (8 weeks old), immunofluorescence staining and in situ hybridization were used to observe the expression and location of Dynamin-1 and miR-140. The luciferase reporter gene vector was constructed and then transfected into 293T cells to determine whether Dnm1 is the target gene of miR-140. Adeno-associated viral vectors carrying miR-140 precursor and blank gene were injected into the cochlea of the mice via circular window membrane. After 14 d, the mRNA expression of miR-140 and Dnm1 was detected by RT-PCR, and Dynamin-1 protein was detected by Western blotting. Results    miR-140 was expressed in the cytoplasm of inner and outer hair cells, and Dynamin-1 was mainly expressed in the cytoplasm of inner hair cells. The luciferase activity assay showed that miR-140 could inhibit the enzyme reporter activity of 3′UTR of Dnm1 (P<0.05). The mRNA expression of miR-140 was significantly increased after transfection of AAV-miR-140, while the mRNA level of Dnm1 and its protein level were decreased significantly (P<0.05). Conclusion    Dnm1 is the target gene of miR-140, and the latter can inhibit the expression of Dynamin-1 in inner ear hair cells, suggesting that it may negatively regulate the process of synaptic vesicle endocytosis of inner hair cells by inhibiting Dynamin, and then affect the transmission of auditory signals.


[1]SCHMID S L, FROLOV V A. Dynamin: functional design of a membrane fission catalyst[J]. Annu Rev Cell Dev Biol, 2011, 27: 79-105. DOI:10.1146/annurevcellbio100109104016.
[2]RAIMONDI A, FERGUSON S M, LOU X, et al. Overlapping role of dynamin isoforms in synaptic vesicle endocytosis[J]. Neuron, 2011, 70(6): 1100-1114. DOI:10.1016/j.neuron.2011.04.031.
[3]BOUMIL R M, LETTS V A, ROBERTS M C, et al. A missense mutation in a highly conserved alternate exon of dynamin1 causes epilepsy in fitful mice[J]. PLoS Genet, 2010, 6(8):e1001046.DOI: 10.1371/journal.pgen.1001046.
[4]蔡婷,陈知己,顾翔,等. Dynamin在不同年龄段小鼠耳蜗中的表达[J]. 中华耳鼻咽喉头颈外科杂志, 2015, 50(7): 583-586.DOI:10.3760/cma.j.issn.16730860.2015.07.012.
CAI T, CHEN Z J, GU X,et al.Expression of dynamin in mouse cochlea of different ages[J]. Chin J Otorhinolaryngol Head Neck Surg, 2015,50(7):583-586. DOI:10.3760/cma.j.issn.16730860.2015.07.012.
[5]USHAKOV K, RUDNICKI A, AVRAHAM K B. MicroRNAs in sensorineural diseases of the ear[J]. Front Mol Neurosci,2013,6:52.DOI:10.3389/fnmol.2013.00052.
[6]HUANG W, LI M D. Nicotine modulates expression of miR140*, which targets the 3′untranslated region of dynamin 1 gene (Dnm1)[J]. Int J Neuropsychopharmacol, 2009, 12(4): 537-546. DOI: 10.1017/S1461145708009528.
[7]PATEL M, HU B H. MicroRNAs in inner ear biology and pathogenesis[J]. Hear Res, 2012, 287(1/2): 6-14.DOI:10.1016/j.heares.2012.03.008.
[8]AKIL O, ROUSE S L, CHAN D K, et al. Surgical method for virally mediated gene delivery to the mouse inner ear through the round window membrane[J]. J Vis Exp, 2015(97):52187.DOI:10.3791/52187.
[9]徐延军,胡吟燕,翟所强,等. 耳后入路圆窗膜显微注射小鼠耳蜗基因转染新途径的研究[J]. 听力学及言语疾病杂志, 2009,17(3): 279-282. DOI:10.3969/j.issn.10067299.2009.03.023.
XU Y J, HU Y Y, ZHAI S Q,et al.The study of a new approach to postauricular microinjection via the round window membrane for cochlear gene transfection in mouse[J].J Audiol Speech Pathol, 2009,17(3):279-282. DOI:10.3969/j.issn.10067299.2009.03.023.
[10]蔡婷,袁伟. 突触内吞关键蛋白Dynamin在小鼠耳蜗内毛细胞中的表达研究[J].中华耳科学杂志, 2014,12(4): 643-646. DOI:10.3969/j.issn.16722922.2014.04.033.
CAI T, YUAN W. Study on the expression of synaptic endocytosis key protein dynamin in mice cochlear inner hair cells[J]. Chin J Otology, 2014,12(4): 643-646.DOI:10.3969/j.issn.16722922.2014.04.033.
[11]ZHANG R, MA J, YAO J. Molecular mechanisms of the cartilagespecific microRNA140 in osteoarthritis[J]. Inflamm Res, 2013, 62(10): 871-877.DOI:10.1007/s0001101306548.
[12]YU L, LU Y, HAN X, et al. microRNA1405p inhibits colorectal cancer invasion and metastasis by targeting ADAMTS5 and IGFBP5[J]. Stem Cell Res Ther, 2016, 7(1): 180. DOI:10.1186/s1328701604385.
[13]YUAN Y, SHEN Y, XUE L,et al. miR140 suppresses tumor growth and metastasis of nonsmall cell lung cancer by targeting insulinlike growth factor 1 receptor[J]. PLoS ONE, 2013, 8(9): e73604.DOI:10.1371/journal.pone.0073604.
[14]GU R, SUN Y F, WU M F,et al. Biological roles of microRNA140 in tumor growth, migration, and metastasis of osteosarcoma in vivo and in vitro[J]. Tumour Biol, 2016, 37(1): 353-360. DOI:10.1007/s1327701538018.
[15]WANG X R, ZHANG X M, ZHEN J,et al. MicroRNA expression in the embryonic mouse inner ear[J]. Neuroreport, 2010, 21(9): 611-617. DOI:10.1097/WNR.0b013e328338864b.
[16]PANG J, XIONG H, LIN P, et al. Activation of miR34a impairs autophagic flux and promotes cochlear cell death via repressing ATG9A: implications for agerelated hearing loss[J]. Cell Death Dis, 2017, 8(10): e3079. DOI:10.1038/cddis.2017.462.
[17]LI Y, LI A, WU J, et al. MiR1825p protects inner ear hair cells from cisplatininduced apoptosis by inhibiting FOXO3a[J]. Cell Death Dis, 2016, 7(9): e2362.DOI:10.1038/cddis.2016.246.
[18]LI Y, PENG A, GE S,et al.miR204 suppresses cochlear spiral ganglion neuron survival in vitro by targeting TMPRSS3[J]. Hear Res, 2014, 314: 60-64.DOI:10.1016/j.heares.2014.05.002.


Last Update: 2018-06-13