Differential neurite outgrowth is required for axon specification by cultured hippocampal neurons

Hideaki Yamamoto, Takanori Demura, Mayu Morita, Gary A. Banker, Takashi Tanii, Shun Nakamura

    Research output: Contribution to journalArticle

    41 Citations (Scopus)

    Abstract

    Formation of an axon is the first morphological evidence of neuronal polarization, visible as a profound outgrowth of the axon compared with sibling neurites. One unsolved question on the mechanism of axon formation is the role of axon outgrowth in axon specification. This question was difficult to assess, because neurons freely extend their neurites in a conventional culture. Here, we leveraged surface nano/micro-modification techniques to fabricate a template substrate for constraining neurite lengths of cultured neurons. Using the template, we asked (i) Do neurons polarize even if all neurites cannot grow sufficiently long? (ii) Would the neurite be fated to become an axon if only one was allowed to grow long? A pattern with symmetrical short paths (20 μm) was used to address the former question, and an asymmetrical pattern with one path extended to 100 μm for the latter. Axon formation was evaluated by tau-1/MAP2 immunostaining and live-cell imaging of constitutively-active kinesin-1. We found that (1) neurons cannot polarize when extension of all neurites is restricted and that (2) when only a single neurite is permitted to grow long, neurons polarize and the longest neurite becomes the axon. These results provide clear evidence that axon outgrowth is required for its specification. Axon formation: no growth, no specification One unsolved question on the mechanism of axon formation by cultured neurons was the role of axon outgrowth in axon specification. Using micropatterned surfaces to extrinsically regulate lengths of individual neurites, we show that differential outgrowth of a future axon is required for axon specification. The results presented here provide clear evidence that an axon cannot be specified without sufficient outgrowth.

    Original languageEnglish
    Pages (from-to)904-910
    Number of pages7
    JournalJournal of Neurochemistry
    Volume123
    Issue number6
    DOIs
    Publication statusPublished - 2012 Dec

    Fingerprint

    Neurons
    Axons
    Neurites
    Specifications
    Neuronal Outgrowth
    Kinesin

    Keywords

    • hippocampal neuron
    • kinesin
    • live-cell imaging
    • micropatterned surface
    • neuronal polarization

    ASJC Scopus subject areas

    • Biochemistry
    • Cellular and Molecular Neuroscience

    Cite this

    Differential neurite outgrowth is required for axon specification by cultured hippocampal neurons. / Yamamoto, Hideaki; Demura, Takanori; Morita, Mayu; Banker, Gary A.; Tanii, Takashi; Nakamura, Shun.

    In: Journal of Neurochemistry, Vol. 123, No. 6, 12.2012, p. 904-910.

    Research output: Contribution to journalArticle

    Yamamoto, Hideaki ; Demura, Takanori ; Morita, Mayu ; Banker, Gary A. ; Tanii, Takashi ; Nakamura, Shun. / Differential neurite outgrowth is required for axon specification by cultured hippocampal neurons. In: Journal of Neurochemistry. 2012 ; Vol. 123, No. 6. pp. 904-910.
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