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Mutations in the β-tubulin gene TUBB2B result in asymmetrical polymicrogyria

Nature Genetics volume 41, pages 746–752 (2009)Cite this article

Abstract

Polymicrogyria is a relatively common but poorly understood defect of cortical development characterized by numerous small gyri and a thick disorganized cortical plate lacking normal lamination. Here we report de novo mutations in a β-tubulin gene, TUBB2B, in four individuals and a 27-gestational-week fetus with bilateral asymmetrical polymicrogyria. Neuropathological examination of the fetus revealed an absence of cortical lamination associated with the presence of ectopic neuronal cells in the white matter and in the leptomeningeal spaces due to breaches in the pial basement membrane. In utero RNAi-based inactivation demonstrates that TUBB2B is required for neuronal migration. We also show that two disease-associated mutations lead to impaired formation of tubulin heterodimers. These observations, together with previous data, show that disruption of microtubule-based processes underlies a large spectrum of neuronal migration disorders that includes not only lissencephaly and pachygyria, but also polymicrogyria malformations.

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Figure 1: Magnetic resonance imaging and histopathology analyses of individuals with TUBB2B mutations.
Figure 2: In utero knockdown of rat Tubb2b expression by RNAi alters neuronal migration in the isocortex.
Figure 3: Various substitutions in TUBB2B result in inefficient α/β tubulin heterodimer formation in vitro. (a) Ribbon presentation illustrating placement of side chains of mutated residues (shown in red) and the E-site guanine nucleotide (shown in orange) within the structure of the β-tubulin polypeptide11,12.
Figure 4: In vitro reactions reveal a lowered affinity of S172P TUBB2B for TBCD.
Figure 5: Loss of function of mutant TUBB2B in vitro, in cultured cells and in vivo.

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Acknowledgements

We thank F. Francis for her helpful comments and critical readings of the manuscript, the subjects and their parents who contributed in this study and all the colleagues who provided clinical and imaging information. We thank R. Guerrini for providing us helpful advice. We are grateful to E. Leguern for allowing K.P. to develop this project and all the members of Cochin Institute genomic platform, Cochin Hospital Cell Bank, and Isabelle Souville for their technical assistance. This work was supported by funding from AP-HP, INSERM, FRM (funding within the frame of the Programme EQUIPEs FRM 2007) and ANR (ANR Neuro 2005, project A05183KS and ANR-06-NEURO-008-01 contract number RPV06055ASA). X.H.J. is supported by a PhD fellowship of the Ministère de l'Enseignement Supérieur et de la Recherche, by a grant for mobility from Université Paris Descartes and an EMBO short-term fellowship (ASTF 66.00-2008) for his work in NYU the Medical Center. K.P. is a post-doctoral researcher supported by FRM (Fondation pour la Recherche Medicale).

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Author notes

  1. Françoise Phan-Dinh-Tuy

    Present address: Present address: UMR-S 839, INSERM, Université Pierre et Marie Curie-Paris 6, Institut du Fer à Moulin, Paris, France.,

  2. Xavier Hubert Jaglin and Karine Poirier: These authors contributed equally to this work.

Authors and Affiliations

  1. Institut Cochin, Université Paris Descartes CNRS (UMR 8104), Paris, France

    Xavier Hubert Jaglin, Karine Poirier, Yoann Saillour, Nadia Bahi-Buisson, Françoise Phan-Dinh-Tuy, Laëtitia Castelnau-Ptakhine & Jamel Chelly

  2. Inserm, U567, Paris, France

    Xavier Hubert Jaglin, Karine Poirier, Yoann Saillour, Nadia Bahi-Buisson, Françoise Phan-Dinh-Tuy, Laëtitia Castelnau-Ptakhine & Jamel Chelly

  3. Plate-forme Post Génomique de l'INMED, INSERM U901, Parc scientifique de Luminy, Marseille, France

    Emmanuelle Buhler

  4. Department of Biochemistry, New York University Medical Center, New York, New York, USA

    Guoling Tian, Xiang Peng Kong & Nicholas Justin Cowan

  5. Département de Pédiatrie, Service de Neurologie Pédiatrique, Hôpital Necker Enfants Malades, AP-HP, Paris, France

    Nadia Bahi-Buisson & Manoelle Kossorotoff

  6. Service d'Anatomie Pathologique, Hôpital Sainte Anne, Paris, France

    Catherine Fallet-Bianco

  7. Institut de Neurobiologie de la Méditerranée, INSERM U901, Parc scientifique de Luminy, Marseille, France

    Pascale Bomont, Carlos Cardoso & Alfonso Represa

  8. Service de génétique médicale, CHU de Rennes-Hôpital Sud, Rennes, France

    Sylvie Odent

  9. Département d'anatomie et cytopathologie, CHU Pontchaillou, Rennes, France

    Philippe Loget

  10. Locatie Juliana Kinderziekenhuis Polikliniek, Den Haag, The Netherlands

    Irina Snoeck

  11. Service de génétique, CHU Hôpital Clémenceau, Avenue Georges Clémenceau, Caen, France

    Ghislaine Plessis

  12. Département de pédiatrie et génétique médicale, CHU Hôpital Morvan, Brest, France

    Philippe Parent

  13. Laboratoire de biochimie et génétique moléculaire, CHU Hôpital Cochin, Paris, France

    Cherif Beldjord

  14. Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK

    Jonathan Flint

  15. Institute of Molecular Pathology, Vienna, Austria

    David Anthony Keays

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  1. Xavier Hubert Jaglin
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Contributions

J.C. coordinated and instigated the study with D.A.K. and J.F. X.H.J. performed biochemical, cellular and in vivo functional studies. N.B.-B., K.P. and C.F.-B. recruited cases and controls. N.B.-B., C.F.-B., S.O., P.L., M.K., I.S., G.P., P.P. and C.B. helped in collecting patients. K.P., D.A.K. and Y.S. screened the subject DNAs and performed the genetic analyses. L.C.-P. performed all DNA extractions from subject samples and coordinated interaction with clinicians. C.F.-B. performed the neuropathological analyses. G.T. and N.J.C. provided reagents and expertise for the biochemical study; X.P.K. helped compute and analyze the structures; C.C., E.B., P.B. and A.R. provided expertise and technical assistance for in utero RNAi analysis. F.P.-D.-T. and K.P. performed the RNA in situ hybridization analysis. X.H.J. and K.P. drafted the manuscript with the help of N.J.C. and J.C.

Corresponding authors

Correspondence to Nicholas Justin Cowan or Jamel Chelly.

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Supplementary Figures 1–12, Supplementary Tables 1 and 2, Supplementary Note and Supplementary Methods (PDF 4891 kb)

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Jaglin, X., Poirier, K., Saillour, Y. et al. Mutations in the β-tubulin gene TUBB2B result in asymmetrical polymicrogyria. Nat Genet 41, 746–752 (2009). https://doi.org/10.1038/ng.380

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