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Mutations in EFHC1 cause juvenile myoclonic epilepsy

Nature Genetics volume 36, pages 842–849 (2004)Cite this article

Abstract

Juvenile myoclonic epilepsy (JME) is the most frequent cause of hereditary grand mal seizures1,2. We previously mapped and narrowed a region associated with JME on chromosome 6p12–p11 (EJM1)3,4,5. Here, we describe a new gene in this region, EFHC1, which encodes a protein with an EF-hand motif. Mutation analyses identified five missense mutations in EFHC1 that cosegregated with epilepsy or EEG polyspike wave in affected members of six unrelated families with JME and did not occur in 382 control individuals. Overexpression of EFHC1 in mouse hippocampal primary culture neurons induced apoptosis that was significantly lowered by the mutations. Apoptosis was specifically suppressed by SNX-482, an antagonist of R-type voltage-dependent Ca2+ channel (Cav2.3). EFHC1 and Cav2.3 immunomaterials overlapped in mouse brain, and EFHC1 coimmunoprecipitated with the Cav2.3 C terminus. In patch-clamp analysis, EFHC1 specifically increased R-type Ca2+ currents that were reversed by the mutations associated with JME.

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Figure 1: Structure and expression of EFHC1.
Figure 2: EFHC1 mutations segregating with epilepsy traits in families with JME.
Figure 3: Activation of R-type VDCC (Cav2.3)-dependent apoptosis by EFHC1 and its reversal by mutations associated with JME.
Figure 4: Colocalization of Efhc1 and Cav2.3.
Figure 5: Potentiation of R-type VDCC (Cav2.3) activity by wild-type and its reversal by mutant EFHC1.
Figure 6: Immunoprecipitation assays of EFHC1 and VDCCs.

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Acknowledgements

We thank the family members for participating in this study; Y. Itsukaichi, K. Yamada, Y. Tsutsumi, K. Shoda, E. Mazaki, A. Nitta, N. Okamura, C. Uchikawa, M. Hishinuma and S.G. Pietsch for their help; and the Research Resources Center of RIKEN Brain Science Institute for DNA sequencing analysis and generation of rabbit polyclonal antibodies. This work was supported in part by a grant from RIKEN Brain Science Institute, Japan. A.V.D.E. is partly supported by a grant from the US National Institutes of Health.

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Author notes

  1. Toshimitsu Suzuki and Kripamoy Aguan: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory for Neurogenetics, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako-shi, 351-0198, Saitama, Japan

    Toshimitsu Suzuki, Kripamoy Aguan, Jun Shi, Tamaki Takeuchi, Ryoji Morita, Subramaniam Ganesh, Yoshihisa Sugimoto & Kazuhiro Yamakawa

  2. Department of Molecular Cytogenetics, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan

    Toshimitsu Suzuki & Johji Inazawa

  3. Epilepsy Genetics/Genomics Laboratories, Comprehensive Epilepsy Program, David Geffen School of Medicine at UCLA and VA GLAHS-West Los Angeles, Room 3405 (127B), Building 500, West Los Angeles DVA Medical Center, 11301 Wilshire Boulevard, Los Angeles, 90073, California, USA

    Antonio V Delgado-Escueta, Marco T Medina, Dongsheng Bai & Julia N Bailey

  4. National Institute of Neurology and Neurosurgery, Mexico City, Mexico

    Maria E Alonso, Adriana Ochoa, Aurelio Jara-Prado, Astrid Rasmussen, Jaime Ramos-Peek, Sergio Cordova & Francisco Rubio-Donnadieu

  5. Center for Integrative Bioscience, The Graduate University for Advanced Studies, Okazaki, Japan

    Yuji Hara, Motohiro Nishida, Tomohiro Numata & Yasuo Mori

  6. Graduate School of Engineering, Kyoto University, Kyoto, Japan

    Yuji Hara, Motohiro Nishida & Yasuo Mori

  7. National Autonomous University, Tegucigalpa, Honduras

    Marco T Medina

  8. Neuropsychiatric Institute, David Geffen School of Medicine at UCLA, Los Angeles, California, USA

    Julia N Bailey

  9. National Epilepsy Center, Shizuoka Medical Institute of Neurological Disorders, Shizuoka, Japan

    Yushi Inoue

  10. Department of Pediatrics, Tokyo Women's Medical University, Tokyo, Japan

    Makiko Osawa & Hirokazu Oguni

  11. Department of Neuropsychiatry, School of Medicine, Hirosaki University, Aomori, Japan

    Sunao Kaneko

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  1. Toshimitsu Suzuki
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Correspondence to Antonio V Delgado-Escueta or Kazuhiro Yamakawa.

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Supplementary information

Supplementary Fig. 1

Amino acid sequence of EFHC1 and evolutionary conservation of amino acids mutated in JME. (PDF 25 kb)

Supplementary Fig. 2

Expression of mouse Efhc1 transcripts and proteins. (PDF 500 kb)

Supplementary Fig. 3

No effect of VDCC antagonists on neurons expressing GFP-controls and binding of EFHC1 mutants to CaV2.3. (PDF 642 kb)

Supplementary Table 1

Modulation of activation and inactivation properties of CaV2.3 by EFH1 proteins. (PDF 397 kb)

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Suzuki, T., Delgado-Escueta, A., Aguan, K. et al. Mutations in EFHC1 cause juvenile myoclonic epilepsy. Nat Genet 36, 842–849 (2004). https://doi.org/10.1038/ng1393

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