Dissection of inner ear neuroepithelium in zebrafish

Abstract

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. 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°C using cryostat, the hematoxylin and eosin staining and immunofluorescence 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. After hematoxylin and eosin staining and immunofluorescence 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 integrity of the sensory maculae could be preserved completely. After immunofluorescence and histochemistry staining, intact epithelia with strong hair cell bundle staining could be seen. The neuroepithelium hair cell examination of zebra fish can be entirely attained by surface preparation of inner ear maculae.