Objective To establish a model by using zebrafish for the inner ear development and disease research,to explore the possibility of complete dissection of inner ear neuroepithelium in zebrafish by surface preparation of inner ear maculae.Methods The inner ear samples of zebra fish were fixed followed by harvesting.After decalcified and dehydrated,the coronal cryosections were made at 3 μm thickness at -23℃ using cryostat,the hematoxylin and eosin staining and immnofluorescence and histochemistry staining were performed,the saccules,utricles and lagenaes were successfully obtained by fine dissection.The sensory maculae were mounted on glass and then were stained.Results After hematoxylin and eosin staining and immnofluorescence and histochemistry staining of cryosections,the sensory maculae of inner ear could be determined by otoliths.On the basis of distinguish of otoliths,the sensory maculae of inner ear could be finely dissected,the overall intergrity of the sensory maculae could be preserved completely.After immnofluorescence and histochemistry staining,intact epithelia with strong hair cell bundle staining could be seen.Conclusions The neuroepithelium hair cell examination of zebra fish can be entirely attained by surface preparation of inner ear maculae.%目的 探索斑马鱼内耳感觉上皮的分离方法,为采用斑马鱼作为内耳发育及疾病研究的动物模型提供技术手段.方法 取4个月龄斑马鱼进行内耳组织冰冻切片,HE染色确定内耳感觉囊斑部位,并行免疫荧光染色观察毛细胞纤毛.在解剖显微镜下将斑马鱼内耳椭圆囊、球囊及听壶完整分离,取出内耳感觉囊斑平铺于载玻片中,进行免疫荧光染色,观察内耳感觉囊斑铺片的形态结构.结果 冰冻切片证实斑马鱼内耳感觉上皮位于囊斑内,并与耳石相对应.以耳石为标志分离出完整的内耳感觉囊斑组织,铺片免疫荧光染色后可观察到内耳感觉上皮形态结构完整,毛细胞纤毛显示清晰.结论 通过内耳感觉囊斑铺片技术可以完整分离出斑马鱼的内耳感觉上皮组织.
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