Abstract
Neuronal migration disorders such as lissencephaly and subcortical band heterotopia are associated with epilepsy and intellectual disability. DCX, PAFAH1B1 and TUBA1A are mutated in these disorders; however, corresponding mouse mutants do not show heterotopic neurons in the neocortex. In contrast, spontaneously arisen HeCo mice display this phenotype, and our study revealed that misplaced apical progenitors contribute to heterotopia formation. While HeCo neurons migrated at the same speed as wild type, abnormally distributed dividing progenitors were found throughout the cortical wall from embryonic day 13. We identified Eml1, encoding a microtubule-associated protein, as the gene mutated in HeCo mice. Full-length transcripts were lacking as a result of a retrotransposon insertion in an intron. Eml1 knockdown mimicked the HeCo progenitor phenotype and reexpression rescued it. We further found EML1 to be mutated in ribbon-like heterotopia in humans. Our data link abnormal spindle orientations, ectopic progenitors and severe heterotopia in mouse and human.
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Acknowledgements
We thank the Inserm Avenir program, the French Agence National de la Recherche (ANR-08-MNP-013 (F.F.), ANR-13-BSV4-0008-01 (F.F.), ANR Blanc 1103-01 (J.C.), projet R11039KK, ANR E-Rare Program, convention 2011-RaARE-012-01) (J.C.), the Fondation Bettencourt Schueller and the Federation pour la recherche sur le cerveau (FRC) for grants awarded to F.F., the FRC Rotary for an equipment grant awarded to the Institut du Fer à Moulin, the Swiss National Science Foundation (SNSF 31003A-135574, SPUM-33CM30-124089 and 33CM30-140332) and the Fondation Gianni Biaggi de Blasys for grants awarded to A.C., and the SNSF (31003A-125379) for grants awarded to E.W. We are particularly grateful to families and clinicians for access to their DNA samples. We also thank B. Barry, C. Walsh and collaborating clinicians for access to certain of their severe heterotopia patients. We thank W. van Ijcken for performing exome sequencing and R. Schot for excellent analysis of exome data of family 3489. We also thank W.B. Dobyns (Seattle Children's Research Institute) for critical review and classification of the MRI of the affected member of family 3489. We are grateful to D. Turner (University of Michigan) and N. Heintz (Rockefeller University) for reagents. We thank the imaging and animal house platforms at the Institut du Fer à Moulin and the Région Ile de France for support, as well as the animal facility and the Cellular Imaging Facility at the Département des Neurosiences Fondamentales. We thank J. Lemarchand, Y. Saillour, D. Derbala, M. Foglio, A. Boland, D. Zelenika, I. Gut, J.P. Hornung, D. Valloton, M. Niquille, C. Devenoges, N. Narboux-Neme, A. Cabrera, E. Bruel-Jungerman, R. Khalaf-Nazzal, X. Jaglin, T.D.H. Iuliano, M. Karababa, N. Magalhaes, M. Nosten-Bertrand, G. Grannec, A. Houllier and I. Moutkine for their contributions to this work. We are grateful to M. Groszer, J.-A. Girault, A. Houdusse, C. Moores, R. Bayliss, A. Fry, S. Cappello and M. Götz for discussions. The group of F.F. is affiliated with the Ecole des Neurosciences de Paris (ENP) and the Bio-Psy laboratory of excellence. The research leading to a part of these results received funding from the European Union Seventh Framework Programme FP7/2007–2013 under the project DESIRE (grant agreement no. 602531), la Fondation pour la Recherche Médicale (FRM, J Chelly–Equipe FRM 2013: DEQ2000326477) la Fondation Jacques Espinasse & Danièle (JED)-Belgique and the Spanish Ministry of Economy and Competitiveness grant (BFU2012-33473) awarded to V.B.
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M.K. and F.P.D.T. carried out HeCo phenotyping and genotyping projects. F.P.D.T. characterized Eml1 at the genomic, cDNA and protein levels. F.P.D.T. and S.B. performed Vero, N2A, progenitor and neuronal cultures and analysis of EML1 and Eml1 (wild type and mutant) in vitro and mouse in situ hybridization studies, aided by K.B. S.B. performed rescue and shRNA in utero electroporations and analyses with help from R.B. M.K. performed the time lapse movies, pictures and reconstructed the movies. C.L. corrected the movie reconstruction and analyzed the films. C.D.J.R. and V.B. performed ferret in situ hybridizations and in utero transplantation analyses. K.P., R. Oegema, G.M.M., N.B.-B., A.G.L.M., P.B. and J.C. screened and characterized patients. K.P. and N.B.-B. followed and screened a patient DNA panel and controls. R. Oegema and G.M.M. performed exome sequencing and mutation confirmation. R. Olaso and J.-F.D. performed transcriptome experiments, genotyping data analysis and database submissions. D.B. aided with microtubule experiments. W.C. performed the second-round mouse genotyping experiment. E.W., A.C. and F.F. directed the study, analyzed the data and wrote the manuscript, greatly helped by M.K., F.P.D.T. and S.B.
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Kielar, M., Tuy, F., Bizzotto, S. et al. Mutations in Eml1 lead to ectopic progenitors and neuronal heterotopia in mouse and human. Nat Neurosci 17, 923–933 (2014). https://doi.org/10.1038/nn.3729
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DOI: https://doi.org/10.1038/nn.3729
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