摘要
目的:在近成年豚鼠耳蜗内携带atoh1/EGFP基因的腺病毒是否能够将基膜上残留的支持细胞转分化为新的毛细胞。方法:选用体重在200～250g的健康花豚鼠12只,将构建同时表达atoh1和EGFP基因的E1/E3区缺失的人类免疫5型腺病毒5μl,通过耳蜗侧壁打孔灌注导入中阶内淋巴系统。其中6只于灌注2周后处死,6只灌注4周后处死,基膜铺片观察EGFP、毛细胞标记物myosinⅦa和Dapi核染色情况。结果:灌注2周处死的6只豚鼠中有2只在基膜外毛细胞外区域发现有散在的细胞核大、细胞体梭形的、表达EGFP的新生细胞。灌注4周的动物中有3只在基膜原外毛细胞位置和外毛细胞外区域发现有相似的同时表达EGFP和Myo-sinⅦa的新生细胞。结论:atoh1基因可以在近成年豚鼠体内将部分支持细胞转分化为毛细胞样细胞。这部分支持细胞位于外毛细胞位置和外毛细胞外区域的基膜上。
Objective：To explore whether the Ad5-atohl/EGFP could transdifferent the supporting ceils into the new hair ceils in young adult guinea pigs cochlea in vivo. Method：Twelve healthy pigmented guinea pigs weigh- ted 200~250 g were included in this experiment. 5 ul of AdS-El/E3 defected-atohl/EGFP were infused into the scala media through a hole made on the lateral wall of the cochlea. Six of the 12 animal were killed 2 weeks after the infusion operation. The others were killed 4 weeks after the operation. The whole mount of the basal mem-branes were directly observed under the fluorescence microscope for the expression of the EGFP（enhance green fluorescent protein） or for the expression of the hair cellspecific marker and nuclear after staining with myosinVIla rabbit polyclonal antibody and Dapi dye. Result： New ceils with big nuclear, ellipse body and expressed with EGFP were found in the region near to the outmost row of the outer hair cells in 2 animal 2 weeks after the infusion. Moreover there were 3 animals with specific morphologic new cells in the location where ever been located by the outer hair cells and the region as 2 weeks animals 4 weeks after the infusion. Those cells were stained by myosinVIIa antibody. Conclusion：Atohl gene could transdifferent some supporting cells in the basal membrane into hair cell like cells in young adult guinea pigs in vivo. These supporting cells locate in the region of outer hair cells and the basal membrane which do not belong to the region of outer hair cells.
出处
《临床耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2012年第10期463-465,共3页
Journal of Clinical Otorhinolaryngology Head And Neck Surgery
基金
上海市科委自然基金项目(No:09ZR1405700)
关键词
腺病毒
atoh1
豚鼠
毛细胞
转分化
adenovirus
atohl
guinea pig
hair cell
transdifferentiation
