Atoh1过表达水平对异位耳蜗毛细胞样细胞的prestin表达及纤毛形态的影响  

Effects of Atoh1 overexpression level on prestin expression and ciliary morphology of ectopic hair-cell-like cells in cochlear explants

作　　者：郑宇 马锐 迟放鲁[1,2,3,4] 赵萌 ZHENG Yu;MA Rui;CHI Fang-lu;ZHAO Meng(Institute of Ear Nose and Throat,Eye Ear Nose and Throat Hospital,Fudan University,Shanghai 200031,China;Department of Otorhinolaryngolgoy,Eye Ear Nose and Throat Hospital,Fudan University,Shanghai 200031,China;Shanghai Clinical Medical Center of Hearing Medicine,Shanghai 200031,China;Key Laboratory of Hearing Medicine,National Health Commission,Shanghai 200031,China;Department of Research Center,Eye Ear Nose and Throat Hospital,Fudan University,Shanghai 200031,China)

机构地区：[1]复旦大学附属眼耳鼻喉科医院耳鼻喉科研究院,上海200031 [2]复旦大学附属眼耳鼻喉科医院耳鼻喉科,上海200031 [3]上海市听觉医学临床中心,上海200031 [4]国家卫生健康委员会听觉医学重点实验室,上海200031 [5]复旦大学附属眼耳鼻喉科医院实验中心,上海200031

出　　处：《复旦学报（医学版）》2020年第2期197-204,共8页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金(81420108010,81271084,81700915)。

摘　　要：目的探讨Atoh1过表达水平对体外培养的耳蜗小上皮嵴区域毛细胞样细胞数量、prestin表达情况及其表面纤毛形态的影响。方法取出生后第1天的大鼠耳蜗中圈进行体外培养,之后采用不同浓度的带有Atoh1基因的腺病毒(含报告基因EGFP)进行转染,检测腺病毒转染后不同时间点体外培养组织的Atoh1 mRNA相对表达量,同时观察小上皮嵴区域绿色荧光蛋白信号强度、毛细胞样细胞数量、毛细胞样细胞的prestin表达情况及其表面纤毛形态。结果带有Atoh1基因的腺病毒转染浓度越高,Atoh1的mRNA表达水平越高,小上皮嵴区域绿色荧光蛋白信号越强,同时该区域生成的毛细胞样细胞数量越多,表达prestin的毛细胞样细胞数量也越多(P<0.05),并且这些细胞表面静纤毛由低到高排列,形态更为规则。结论Atoh1过表达水平与诱导生成的能够表达prestin的异位耳蜗毛细胞样细胞数量正相关,并且Atoh1过表达水平越高,这些细胞的表面纤毛形态越规则。Objective To investigate the effects of Atoh1 overexpression level on the number,prestin expression and ciliary morphology of ectopic hair-cell-like cells in the lesser epithelium ridge of cochlear explants.Methods The middle turn of one-day-old rats’cochlea was cultured in vitro,followed by transfection of different concentrations of adenovirus encoding Atoh1 and reporter gene EGFP.Then we examined relative mRNA expression level of Atoh1 of the explants and the intensity of green fluorescent protein in the lesser epithelial ridge at different time points;we also examined the number,prestin expression and ciliary morphology of ectopic hair-cell-like cells in the lesser epithelium ridge.Results The higher the concentration of adenovirus encoding Atoh1 led to higher Atoh1 mRNA expression level in cochlear explants,and gave rise to greater number of hair cell-like cells and stronger intensity of green fluorescent protein in the lesser epithelium ridge as well as greater the number of hair cell-like cells with expression of prestin(P<0.05);besides,these cells possessed a more regular cilium with stereocilium lining up from shortest to longest according to its length.Conclusion The overexpression level of Atoh1 is positively correlated with the number of ectopic cochlear hair cell-like cells with expression of prestin and the higher expression level of Atoh1,the more organized cilium of these newly generated hair cell-like cells.

关 键 词：Atoh1 小上皮嵴 PRESTIN 纤毛 毛细胞样细胞 

分 类 号：R764[医药卫生—耳鼻咽喉科]