The transmembrane semaphorin protein Sema6A is broadly expressed in the developing nervous system. Sema6A repels several classes of developing axons in vitro and contributes to thalamocortical axon guidance in vivo. Here we show that during cerebellum development, Sema6A is selectively expressed by postmitotic granule cells during their tangential migration in the deep external granule cell layer, but not during their radial migration. In Sema6A-deficient mice, many granule cells remain ectopic in the molecular layer where they differentiate and are contacted by mossy fibers. The analysis of ectopic granule cell morphology in Sema6a−/− mice, and of granule cell migration and neurite outgrowth in cerebellar explants, suggests that Sema6A controls the initiation of granule cell radial migration, probably through a modulation of nuclear and/or soma translocation. Finally, the analysis of mouse chimeras suggests that this function of Sema6A is primarily non-cell-autonomous.
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In the version of the supplementary information originally attached to this article at the time of its publication online, the legends to supplementary figures were missing. This error has been corrected.
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We thank C. Sotelo for his constant support and comments on the manuscript; H. Sakagami and H. Kondo (Tohoku University) for anti-β isoform of CaMKIV antibody; R. Hawkes (University of Calgary, Canada) for Zebrin II Q113 antibody; D. Karagogeos (University of Crete, Greece) for TAG-1 antibody; F. Qiu (UMDNJ, Piscataway, USA) for Barhl1 cDNA; M. Wassef (ENS, Paris, France) for Math1 cDNA; K. Skurka and D. Rottkamp for help in localizing the insertion site in the Sema6a gene trap allele; R. Schwartzmann and V. Georget (Service d'Imagerie IFR83, Université Paris 6, France) for their help with confocal and videomicroscopy studies; and M. Wassef, C. Lebrand, C. Métin and J.P. Baudoin for discussions. A.C. is supported by the Fondation pour la Recherche sur le Cerveau (FRC), the Schlumberger Foundation and the Association pour la Recherche sur le Cancer (ARC). K.J.M is a Science Foundation Ireland (SFI) Investigator. This work was supported by SFI grant 01/F.1/B006 to K.J.M; and grants from the 21st Century COE Program, and from the Core Research for Evolutional Science and Technology (CREST) of the Japan Science and Technology Agency (JST) to H.F.
The authors declare no competing financial interests.
BrdU pulse labelling. (PDF 2325 kb)
Distribution of granule cell precursors. (PDF 3019 kb)
Granule cell neurite outgrowth. (PDF 698 kb)
Model of Sema6A function. (PDF 395 kb)
Time-lapse video 1 of migrating wild-type neurons. Migration of wild-type granule cells from an EGL microexplant after 24 h in culture. The explant is on the left of the frame. Granule cells migrate away from the explant. They also extend long processes bearing dynamic growth cones. (MOV 1962 kb)
Time-lapse video 2 of migrating wild-type neurons. Wild-type EGL microexplant cultured for 24 h. The explant is on the top. The movement of the soma/nucleus is saltatory and preceded by the appearance of an elongated swelling just ahead of the nucleus. (MOV 1018 kb)
Time-lapse video 3 of migrating Sema6A−/− neurons. Sema6A−/− EGL microexplant after 24 h in culture. The explant is on the left of the frame. Many granule cells remains stationary, oscillating, and some move backward towards the explant. However the cells extend long neurites as active as wild-type ones. (MOV 1251 kb)
Time-lapse video 4 of migrating Sema6A−/− neurons. The explant is located on the left of the frame. The cell body of most Sema6A−/− granule cells does not move forward and one in the center of the frame move back toward the explant. (MOV 1303 kb)
Time-lapse video 5 of migrating Sema6A−/− neurons. The explant is on the left of the frame. Illustration of two migrating Sema6A−/− granule cells. One in the bottom moves rearward toward the explant, while the second one migrate in the correct direction. (MOV 754 kb)
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Kerjan, G., Dolan, J., Haumaitre, C. et al. The transmembrane semaphorin Sema6A controls cerebellar granule cell migration. Nat Neurosci 8, 1516–1524 (2005). https://doi.org/10.1038/nn1555
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