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

郑宇, 马锐, 迟放鲁, 赵萌

复旦学报(医学版) ›› 2020, Vol. 47 ›› Issue (02) : 197-204.

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复旦学报(医学版) ›› 2020, Vol. 47 ›› Issue (02) : 197-204. DOI: 10.3969/j.issn.1672-8467.2020.02.010
论著

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

  • 郑宇1,2,3,4, 马锐1,3,4,5, 迟放鲁1,2,3,4, 赵萌1,2,3,4
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Effects of Atoh1 overexpression level on prestin expression and ciliary morphology of ectopic hair-cell-like cells in cochlear explants

  • ZHENG Yu1,2,3,4, MA Rui1,3,4,5, CHI Fang-lu1,2,3,4, ZHAO Meng1,2,3,4
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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。

Abstract

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

Key words

Atoh1 / lesser epithelial ridge / prestin / cilium / hair-cell-like cells

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导出引用
郑宇, 马锐, 迟放鲁, 赵萌. 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[J]. 复旦学报(医学版), 2020, 47(02): 197-204 https://doi.org/10.3969/j.issn.1672-8467.2020.02.010
ZHENG Yu, MA Rui, CHI Fang-lu, ZHAO Meng. Effects of Atoh1 overexpression level on prestin expression and ciliary morphology of ectopic hair-cell-like cells in cochlear explants[J]. Fudan University Journal of Medical Sciences, 2020, 47(02): 197-204 https://doi.org/10.3969/j.issn.1672-8467.2020.02.010
中图分类号: R764   

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基金

国家自然科学基金(81420108010,81271084,81700915)
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