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Plexin-A2 and its ligand, Sema6A, control nucleus-centrosome coupling in migrating granule cells

Nature Neuroscience volume 11, pages 440–449 (2008)Cite this article

Abstract

During their migration, cerebellar granule cells switch from a tangential to a radial mode of migration. We have previously demonstrated that this involves the transmembrane semaphorin Sema6A. We show here that plexin-A2 is the receptor that controls Sema6A function in migrating granule cells. In plexin-A2–deficient (Plxna2−/−) mice, which were generated by homologous recombination, many granule cells remained in the molecular layer, as we saw in Sema6a mutants. A similar phenotype was observed in mutant mice that were generated by mutagenesis with N-ethyl-N-nitrosourea and had a single amino-acid substitution in the semaphorin domain of plexin-A2. We found that this mutation abolished the ability of Sema6A to bind to plexin-A2. Mouse chimera studies further suggested that plexin-A2 acts in a cell-autonomous manner. We also provide genetic evidence for a ligand-receptor relationship between Sema6A and plexin-A2 in this system. Using time-lapse video microscopy, we found that centrosome-nucleus coupling and coordinated motility were strongly perturbed in Sema6a−/− and Plxna2−/− granule cells. This suggests that semaphorin-plexin signaling modulates cell migration by controlling centrosome positioning.

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Figure 1: Plexin-A2 expression and function in migrating granule cells.
Figure 2: Abnormal in vitro migration of Plxna2−/− granule cells.
Figure 3: Analysis of NMF454 ENU-Plxna2 mutants.
Figure 4: Cell-autonomous function of plexin-A2 in migrating granule cells.
Figure 5: Abnormal nucleus-centrosome coupling in Sema6a and Plxna2 mutants.
Figure 6: Analysis of nucleus-centrosome and nucleus-Golgi spacing in Sema6a and Plxna2 mutants.
Figure 7: Cytoskeleton and Golgi apparatus in Sema6a and Plxna2 mutant granule cells.
Figure 8: Nucleus-centrosome coupling in vivo in ectopic granule cells.

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Acknowledgements

We thank H. Sakagami and H. Kondo for antibody to the β isoform of CaMKIV, M. Okabe for GFP transgenic mice, M. Bornens for centrin-GFP plasmid and antibodies to centrin, and L. Beverly-Staggs and L. Marquis for technical assistance with the NMF454 mice, the production of which was supported by the US National Institutes of Health (NS041215 and NS35900). We also thank Y.E. Jones and R. Robinson for their help with the analysis of plexin-A2 structure and R. Schwartzmann for help with confocal microscopy studies. A.C. and G.K. were supported by the Association pour la Recherche sur le Cancer, the Fondation pour la Recherche Médicale (programme équipe FRM) and the Agence Nationale pour la Recherche (ANR Neurosciences). H.F. was supported by the 21st Century Centers of Excellence Program and Grants-in-Aid for Scientific Research Japan. K.J.M. was supported by a Science Foundation Ireland grant (01/F.1/B006). S.L.A. is an investigator of the Howard Hughes Medical Institute. This work was also supported by grants from CREST (F.S.) of the Japanese Science and Technology Agency. J.R. is recipient of a fellowship from the Région Ile-de-France.

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  1. Alain Chédotal

    Present address: Present address: Institut de la Vision, INSERM, UMR_S592, 17 rue Moreau, Paris F-75012, France.,

Authors and Affiliations

  1. Centre National de la Recherche Scientifique, UMR7102, 9 Quai Saint-Bernard, Paris, F-75005, France

    Julie Renaud, Géraldine Kerjan, Yvrick Zagar, Coralie Fouquet & Alain Chédotal

  2. Université Pierre et Marie Curie–Paris 6, UMR7102, Paris, F-75005, France

    Julie Renaud, Géraldine Kerjan, Yvrick Zagar, Virginie Georget, Coralie Fouquet & Alain Chédotal

  3. Division of Biological Science, Nagoya University Graduate School of Science, Nagoya, 468-8602, Japan

    Itsuko Sumita, Kazunori Suda & Hajime Fujisawa

  4. Institut Fédératif de Biologie Intégrative, 9 Quai Saint-Bernard, Paris, F-75005, France

    Virginie Georget

  5. Howard Hughes Medical Institute, Jackson Laboratory, 600 Main Street, Bar Harbor, 04609, Maine, USA

    Doyeun Kim & Susan L Ackerman

  6. Laboratory of Neurobiology and Behavioral Genetics, National Institute for Physiological Science, Myodaiji, Okazaki, 444-8585, Japan

    Makoto Sanbo

  7. Division of Developmental Genetics, National Institute of Genetics, Mishima, 411-8540, Japan

    Fumikazu Suto

  8. Smurfit Institute of Genetics, Trinity College Dublin, Dublin, 2, Ireland

    Kevin J Mitchell

  9. AP-HP, Groupe Hospitalier Pitié-Salpétrière, Fédération de Neurologie, 91 Boulevard de l'Hôpital, Paris, F-75013, France

    Alain Chédotal

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Contributions

J.R., G.K. and I.S. carried out the in vivo phenotypic analyses of knockout mice and the expression studies. J.R. performed the in vitro experiments. Y.Z. carried out the biochemical studies. C.F. generated the plexin-A2A396E construct and did the binding experiments. V.G. and J.R. performed the time-lapse microscopy and analyzed the confocal images. F.S. and H.F. obtained the Plxna2 knockout and antibodies. K.J.M. provided the Sema6a knockout and helped in the writing of the manuscript. D.K. and S.L.A. generated the NMF454 ENU-mutant and mapped the mutation to the Plxna2 locus. K.S. and M.S. generated the mouse chimeras. A.C. and H.F. designed the study, prepared the figures and wrote the core of the manuscript.

Corresponding author

Correspondence to Alain Chédotal.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–5 (PDF 887 kb)

Supplementary Video 1

Time-lapse video 1 of migrating wild-type granule cells expressing centrin 1–GFP. The explant is on the left of the frame. (MOV 211 kb)

Supplementary Video 2

Time-lapse video 2 of migrating wild-type granule cells expressing centrin 1–GFP. The explant is on the left of the frame. (MOV 72 kb)

Supplementary Video 3

Time-lapse video 3 of migrating Plxna2−/− granule cells expressing centrin 1–GFP. The explant is on the left of the frame. (MOV 116 kb)

Supplementary Video 4

Time-lapse video 4 of migrating Plxna2−/− granule cells expressing centrin 1–GFP. The explant is on the left of the frame. (MOV 188 kb)

Supplementary Video 5

Time-lapse video 5 of migrating Sema6a−/− granule cells expressing centrin 1–GFP. The explant is on the left of the frame. (MOV 131 kb)

Supplementary Video 6

Time-lapse video 6 of migrating Sema6a−/− granule cells expressing centrin 1–GFP. The explant is on the left of the frame. (MOV 129 kb)

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Renaud, J., Kerjan, G., Sumita, I. et al. Plexin-A2 and its ligand, Sema6A, control nucleus-centrosome coupling in migrating granule cells. Nat Neurosci 11, 440–449 (2008). https://doi.org/10.1038/nn2064

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