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UNC-33 (CRMP) and ankyrin organize microtubules and localize kinesin to polarize axon-dendrite sorting

Nature Neuroscience volume 15, pages 48–56 (2012)Cite this article

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Abstract

The polarized distribution of neuronal proteins to axons and dendrites relies on microtubule-binding proteins such as CRMP, directed motors such as the kinesin UNC-104 (Kif1A) and diffusion barriers such as ankyrin. The causative relationships among these molecules are unknown. We show here that Caenorhabditis elegans CRMP (UNC-33) acts early in neuronal development, together with ankyrin (UNC-44), to organize microtubule asymmetry and axon-dendrite sorting. In unc-33 and unc-44 mutants, axonal proteins were mislocalized to dendrites and vice versa, suggesting bidirectional failures of axon-dendrite identity. unc-44 directed UNC-33 localization to axons, where it was enriched in a region that resembled the axon initial segment. unc-33 and unc-44 were both required to establish the asymmetric dynamics of axonal and dendritic microtubules; in their absence, microtubules were disorganized, the axonal kinesin UNC-104 invaded dendrites, and inappropriate UNC-104 activity randomized axonal protein sorting. We suggest that UNC-44 and UNC-33 direct polarized sorting through their global effects on neuronal microtubule organization.

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Figure 1: unc-33 mutants mislocalize presynaptic proteins to dendrites.
Figure 2: UNC-33L functions in PVD during the establishment of polarity.
Figure 3: UNC-33L is enriched in PVD axons.
Figure 4: unc-104 kinesin mislocalizes presynaptic proteins to dendrites in unc-33 mutants.
Figure 5: UNC-104 is mislocalized to dendrites in unc-33 mutants.
Figure 6: A sensory chemoreceptor protein is mislocalized to axons in unc-33 mutants.
Figure 7: unc-44/ankyrin mutations disrupt polarized protein sorting and UNC-33L localization.
Figure 8: Microtubule defects in unc-33 and unc-44 mutants.

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Acknowledgements

We thank C. Ghenoiu, P. Nurse, Y. Saheki, S. Shaham, M. Tsunozaki, M. Heiman, A. Kelly, T. Starich and members of our labs for advice and comments on the manuscript, C. Janke (Institut Curie) and B. Edde (Universite Pierre et Marie Curie) for antibodies, and Y. Saheki (Rockefeller University) and the Caenorhabditis Genetics Center for strains. C.I.B. and K.S. are Investigators of the Howard Hughes Medical Institute. This work was supported by the Howard Hughes Medical Institute.

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Authors and Affiliations

  1. Laboratory of Neural Circuits and Behavior, Howard Hughes Medical Institute, The Rockefeller University, New York, New York, USA

    Tapan A Maniar, Miriam Kaplan & Cornelia I Bargmann

  2. Department of Biology, Howard Hughes Medical Institute, Stanford University, California, USA

    George J Wang & Kang Shen

  3. Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota, USA

    Li Wei & Jocelyn E Shaw

  4. National Centre for Biological Sciences – Tata Institute of Fundamental Research, Bangalore, India

    Sandhya P Koushika

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Contributions

T.A.M. designed, conducted and interpreted most experiments and wrote the paper; M.K., G.J.W., K.S., L.W. and J.E.S. conducted and interpreted individual experiments with unc-33 and unc-44 mutants; S.P.K. generated the UNC-104 antibody and helped design transport experiments; C.I.B. designed and interpreted experiments and wrote the paper.

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Correspondence to Cornelia I Bargmann.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–7 (PDF 2124 kb)

Supplementary Video 1

Movement of EBP-2::GFP puncta in axons of wild type sensory neurons. Cell bodies are to the right. Displayed at 10x speed. Scale bar, 5 μm. (MOV 108 kb)

Supplementary Video 2

Movement of EBP-2::GFP puncta in proximal dendrites of wild type sensory neurons. Distal dendrites are to the left; cell bodies are to the right. Displayed at 10x speed. Scale bar, 5 μm. (MOV 561 kb)

Supplementary Video 3

Movement of EBP-2::GFP puncta in distal dendrites of wild type sensory neurons. Cilia are to the left; cell bodies are to the right (neither is visible). Displayed at 10x speed. Scale bar, 5 μm. (MOV 159 kb)

Supplementary Video 4

Movement of EBP-2::GFP puncta in axons of unc-33(mn407) sensory neurons. Cell bodies are to the right. Displayed at 10x speed. Scale bar, 5 μm. (MOV 183 kb)

Supplementary Video 5

Movement of EBP-2::GFP puncta in proximal dendrites of unc-33(mn407) sensory neurons. Distal dendrites are to the left; cell bodies are to the right. Displayed at 10x speed. Scale bar, 5 μm. (MOV 200 kb)

Supplementary Video 6

Movement of EBP-2::GFP puncta in distal dendrites of unc-33(mn407) sensory neurons. Cilia are to the left; cell bodies are to the right (neither is visible). Displayed at 10x speed. Scale bar, 5 μm. (MOV 96 kb)

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Maniar, T., Kaplan, M., Wang, G. et al. UNC-33 (CRMP) and ankyrin organize microtubules and localize kinesin to polarize axon-dendrite sorting. Nat Neurosci 15, 48–56 (2012). https://doi.org/10.1038/nn.2970

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