Turnover of fluorescently labelled tubulin and actin in the axon

Abstract

THE cytoskeleton has an important role in the generation and maintenance of the structure of the axon^1,2. Microtubules, neurofilaments and actin, together with various kinds of associated proteins, form highly organized dynamic cytoskeletal structures^2–4. Because tubulin and actin molecules are essential cytoskeletal components² and are transported down the axon^5,6, it is important to understand their dynamic behaviour within the axon. Although previous pulse-labelling studies have indicated that the axonal cytoskeleton is a static complex travelling down the axon⁶, this view has been challenged by the results of several recent experiments^7–14. We have now addressed this question by analysing the recovery of fluorescence after photobleaching fluorescent analogues of tubulin and actin in the axons of cultured neurons. We did not observe movement or spreading of bleached zones along the axon, both in neurons injected with fluorescein-labelled tubulin and actin. All bleached zones recovered their fluorescence gradually, however, indicating that microtubules and actin filaments are not static polymers moving forward within the axon, but are dynamic structures that continue to assemble along the length of the axon.