In Vitro Preparation of Newt Inner Ear Sensory Epithelia as a Model for Repair and Regeneration

Abstract

The sensory “hair” cells of the inner ear transform sound energy into electrical signals, but are readily lost through aging, excessive noise, and ototoxic agents. The newt provides an excellent model in which to explore regeneration and whilst loss of hair cells from inner ear epithelia does not require whole organ regeneration, new hair cells are generated from differentiated supporting cells that transdifferentiate without an intervening mitotic event. Here we describe the methods for maintaining the sensory epithelia in long term culture; for the use of the aminoglycoside, gentamicin, to kill the hair cells; and for the examination of the tissue by electron microscopy or fluorescence microscopy. Demembranation of the epithelium reveals the underlying ultrastructure of the tissue for examination by scanning electron microscopy (SEM) and is a technique that can be utilized with immunogold labelling.