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CRMP-2 regulates polarized Numb-mediated endocytosis for axon growth

Nature Cell Biology volume 5, pages 819–826 (2003)Cite this article

Abstract

Axon growth during neural development is highly dependent on both cytoskeletal re-organization and polarized membrane trafficking. Previously, we demonstrated that collapsin response mediator protein-2 (CRMP-2) is critical for specifying axon/dendrite fate and axon growth in cultured hippocampal neurons, possibly by interacting with tubulin heterodimers and promoting microtubule assembly. Here, we identify Numb as a CRMP-2-interacting protein. Numb has been shown to interact with α-adaptin and to be involved in endocytosis. We found that Numb was associated with L1, a neuronal cell adhesion molecule that is endocytosed and recycled at the growth cone, where CRMP-2 and Numb were colocalized. Furthermore, expression of dominant-negative CRMP-2 mutants or knockdown of CRMP-2 message with small-interfering (si) RNA inhibited endocytosis of L1 at axonal growth cones and suppressed axon growth. These results suggest that in addition to regulating microtubule assembly, CRMP-2 is involved in polarized Numb-mediated endocytosis of proteins such as L1.

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Figure 1: Interaction of CRMP-2 with Numb in vivo and in vitro.
Figure 2: Localization of Numb and CRMP-2 in hippocampal neurons.
Figure 3: Association of Numb with L1.
Figure 4: Inhibition of L1 endocytosis at the axonal growth cones.
Figure 5: CRMP-2 is required for axon growth and L1 endocytosis.

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Acknowledgements

We thank Y. Gu, Y. Ihara, A. Craig, M. Nakafuku, A. Tokunaga, H. Okano and V. Lemmon for their kind gifts of materials. We also thank M. Amano, M. Fukata (Nagoya University), H. Qadota and N. Inagaki (Nara Institute of Science and Technology) for their helpful discussions, T. Kimura, T. Shiromizu, H. Watanabe and M. Yoshizaki for preparing some materials and technical assistance, and T. Ishii for secretarial assistance. This research was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan and the organization for Pharmaceutical Safety and Research.

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Authors and Affiliations

  1. Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, 65 Tsurumai, Showa, Nagoya, 466-8550, Aichi, Japan

    Takashi Nishimura, Yuko Fukata, Katsuhiro Kato, Tomoya Yamaguchi & Kozo Kaibuchi

  2. Research Center for Emerging Infectious Disease, Osaka University, 3-1 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Yoshiharu Matsuura

  3. Developmental Brain Science Group, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako, 351-0198, Saitama, Japan

    Hiroyuki Kamiguchi

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  1. Takashi Nishimura
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Correspondence to Kozo Kaibuchi.

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Nishimura, T., Fukata, Y., Kato, K. et al. CRMP-2 regulates polarized Numb-mediated endocytosis for axon growth. Nat Cell Biol 5, 819–826 (2003). https://doi.org/10.1038/ncb1039

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