Transient expression of neurofilament protein during hair cell development in the mouse cochlea

A polyclonal antiserum raised against the 145 kDa neurofilament protein (NFM) has been used to study the distribution of neurofilaments both in organotypic cultures of the early postnatal mouse cochlea and during development of the mouse cochlea in vivo. In the cultures, both the inner hair cells and the outer hair cells are stained by the antibodies, as well as the innervating afferent fibres from the spiral ganglion. In cultures denervated at the time of preparation, neurofilament positive hair cells can still be detected after 7 days in vitro. NFM can also be detected by immunoblotting in such denervated cultures. Characteristic 10 nm diameter, cytoplasmic filaments can also be observed in cultured hair cells using transmission electron microscopy. Immunostaining of cryosections prepared from cochleas at embryonic days 17 and 19, and days 1, 2, 5, 10 and 21 post-partum reveals that hair cells transiently express NFM during their development in vivo. Expression of NFM in hair cells is first detected at embryonic day 19 in the basal region of the cochlea and, by 2 days post-partum, neurofilament positive hair cells are found throughout the entire length of the cochlea. By 10 days post-partum, staining of hair cells begins to diminish and, by 21 days post-partum, NFM can no longer be detected in hair cells.