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miR-140在内耳毛细胞中的表达及其对突触蛋白Dynamin-1的调控作用
Expression of miR-140 in mouse inner ear hair cells and its regulation on synaptic protein Dynamin-1
- 第三军医大学学报 2018年40卷第11期 页码：966-972
- 作者机构：
- 作者简介：
- 基金信息：国家自然科学基金面上项目(81470694);国家自然科学基金面上项目(81271080);西南医院院管课题(JCZD-39)
DOI：10.16016/j.1000-5404.201801142
中图分类号：收稿：2018-01-17，
纸质出版：2018-06
- 稿件说明：
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唐小林, 李洋, 余菲, 等. miR-140在内耳毛细胞中的表达及其对突触蛋白Dynamin-1的调控作用[J]. 第三军医大学学报, 2018,40(11):966-972.
Xiaolin TANG, Yang LI, Fei YU, et al. Expression of miR-140 in mouse inner ear hair cells and its regulation on synaptic protein Dynamin-1[J]. Journal of Third Military Medical University, 2018, 40(11): 966-972.
唐小林, 李洋, 余菲, 等. miR-140在内耳毛细胞中的表达及其对突触蛋白Dynamin-1的调控作用[J]. 第三军医大学学报, 2018,40(11):966-972. DOI： 10.16016/j.1000-5404.201801142.
Xiaolin TANG, Yang LI, Fei YU, et al. Expression of miR-140 in mouse inner ear hair cells and its regulation on synaptic protein Dynamin-1[J]. Journal of Third Military Medical University, 2018, 40(11): 966-972. DOI： 10.16016/j.1000-5404.201801142.
摘要
Abstract
Objective
2
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
2
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
2
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
2
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.
关键词
Keywords
references
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