Association of ezrin isoforms with the neuronal cytoskeleton

We are studying the changes in the organization of the cytoskeleton which accompany expression of differentiated neuronal morphology. Of particular interest is the elaboration of growth cones, the motile domains of the neuronal plasma membrane, and the cytoskeletal structures that underlie them. A candidate for a component of the growth cone cytoskeleton of cultured hippocampal neurons is the antigen recognized by the monoclonal antibody, 13H9 (Birgbauer and Solomon, J Cell Biol 109:1609-1620, 1989; Goslin et al., J Cell Biol 109:1621-1631, 1989). That antibody binds strongly to growth cones, but barely stains neurites. The characterization of the antigen, both biochemical and microscopic, suggests that it may interact with microfilaments and microtubules. We have established that 13H9 recognizes a subset of the isoforms of ezrin (unpublished results). Here, we describe the properties and localization of ezrin isoforms in differentiating neuronal cells, using two in vitro systems and developing spinal cord. In embryonal carcinoma cells, both the abundance of ezrin and the proportion of ezrin associated with the cytoskeletal fraction increase upon induction of neuronal differentiation with retinoic acid. In the neuronal cells within such cultures, the 13H9-positive forms of ezrin are enriched in the growth cone, while the bulk of ezrin identified by a polyclonal antibody shows no specific localization. In mouse DRG neurons, 13H9 staining is asymmetrically distributed along the edges of the complex growth cones of these cells. Staining of developing spinal cord in rat embryos also demonstrates that the 13H9-positive forms of ezrin do not colocalize with the majority of ezrin.(ABSTRACT TRUNCATED AT 250 WORDS)