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Neurogenin 2 controls cortical neuron migration through regulation of Rnd2

Abstract

Motility is a universal property of newly generated neurons. How cell migration is coordinately regulated with other aspects of neuron production is not well understood. Here we show that the proneural protein neurogenin 2 (Neurog2), which controls neurogenesis in the embryonic cerebral cortex1,2, directly induces the expression of the small GTP-binding protein Rnd2 (ref. 3) in newly generated mouse cortical neurons before they initiate migration. Rnd2 silencing leads to a defect in radial migration of cortical neurons similar to that observed when the Neurog2 gene is deleted. Remarkably, restoring Rnd2 expression in Neurog2-mutant neurons is sufficient to rescue their ability to migrate. Our results identify Rnd2 as a novel essential regulator of neuronal migration in the cerebral cortex and demonstrate that Rnd2 is a major effector of Neurog2 function in the promotion of migration. Thus, a proneural protein controls the complex cellular behaviour of cell migration through a remarkably direct pathway involving the transcriptional activation of a small GTP-binding protein.

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Figure 1: Rnd2 expression in the embryonic cerebral cortex requires Neurog2.
Figure 2: Rnd2 silencing blocks the radial migration of cortical projection neurons and alters their morphology.
Figure 3: Expression of Rnd2 rescues the radial migration defect of Neurog2 -mutant cortical neurons.
Figure 4: Neurog2 activates Rnd2 expression in the cerebral cortex by direct binding to a 3′ enhancer element.

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Acknowledgements

We are grateful to S. Wood for technical assistance in generating transgenic embryos. We thank members of the Guillemot laboratory for suggestions on the manuscript, and F. Polleux (University of North Carolina) for the pNeuroD1 plasmid and D. Anderson (California Institute of Technology) for the anti-Neurog2 antibody. J.I.-T.H. is supported by an Australian CJ Martin Fellowship (grant number 310616) and a Medical Research Council career development fellowship (UK), L.N. and H.W. by postdoctoral fellowships awarded by the European Molecular Biology Organization, and D.S.C. and C.Z. by Medical Research Council career development fellowships (UK). This work was supported by grants from the European Commission 6th Framework Programme for Research and Technological Development to F.G. and by institutional funds from the Medical Research Council (UK).

Author Contributions J.I.-T.H. and F.G. conceived all experiments, analysed data and wrote the manuscript. Together with J.I-T.H., L.N. participated in ex vivo electroporation studies, H.W. conducted in utero electroporation experiments, C.Z. performed whole-embryo culture experiments, and D.S.C., D.S.-K. and J.-M.M. performed chromatin immunoprecipitation experiments. O.A. conducted the microarray screen, and F.B. and R.H. contributed to luciferase assays and immunostaining. All authors discussed results from experiments and commented on the manuscript.

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Correspondence to François Guillemot.

Supplementary information

Supplementary Information 1

This file contains Supplementary Figures 1 to 8 with Legends. (PDF 2881 kb)

Supplementary information 2

This file contains Supplementary Figures 9 to 16 for the manuscript. This file also contains Supplementary Table 1 which displays all statistics generated for the experiments herein, as well as legends to Supplementary Movies 1 to 7. (PDF 1240 kb)

Supplementary Movie 1

This file contains Supplementary Movie 1. This movie shows a control multipolar neuron undergoing migration within the intermediate zone. (AVI 2662 kb)

Supplementary Movie 2

This file contains Supplementary Movie 2. This movie shows an Rnd2siRNA-treated multipolar neuron undergoing migration within the intermediate zone. (AVI 3425 kb)

Supplementary Movie 3

This file contains Supplementary Movie 3. This movie shows a control uni/bipolar neuron undergoing migration within the cortical plate. (AVI 3143 kb)

Supplementary Movie 4

This file contains Supplementary Movie 4. This movie shows a Rnd2siRNA-treated uni/bipolar neuron undergoing migration within the cortical plate. (AVI 2756 kb)

Supplementary Movie 5

This file contains Supplementary Movie 5. This movie shows a neuron within the intermediate zone electroporated with the high-expression construct iZ/Rnd2. The neuron lacks neurite outgrowth and appear round. (AVI 2582 kb)

Supplementary Movie 6

This file contains Supplementary Movie 6. This movie shows neurons within the cortical plate electroporated with the high-expression construct iZ/Rnd2. The neuron exhibits a migration defect as well as stabilized leading processes. (AVI 3114 kb)

Supplementary Movie 7

This file contains Supplementary Movie 7. This movie shows neurons within the cortical plate electroporated with the low-expression construct pNeuroD1-Rnd2. The neuron exhibits a migration defect, as well as a stabilized leading processes and excess branching. (AVI 3679 kb)

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Heng, JT., Nguyen, L., Castro, D. et al. Neurogenin 2 controls cortical neuron migration through regulation of Rnd2. Nature 455, 114–118 (2008). https://doi.org/10.1038/nature07198

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