Monogenic neurodevelopmental disorders provide key insights into the pathogenesis of disease and help us understand how specific genes control the development of the human brain. Timothy syndrome is caused by a missense mutation in the L-type calcium channel Cav1.2 that is associated with developmental delay and autism1. We generated cortical neuronal precursor cells and neurons from induced pluripotent stem cells derived from individuals with Timothy syndrome. Cells from these individuals have defects in calcium (Ca2+) signaling and activity-dependent gene expression. They also show abnormalities in differentiation, including decreased expression of genes that are expressed in lower cortical layers and in callosal projection neurons. In addition, neurons derived from individuals with Timothy syndrome show abnormal expression of tyrosine hydroxylase and increased production of norepinephrine and dopamine. This phenotype can be reversed by treatment with roscovitine, a cyclin-dependent kinase inhibitor and atypical L-type–channel blocker2,3,4. These findings provide strong evidence that Cav1.2 regulates the differentiation of cortical neurons in humans and offer new insights into the causes of autism in individuals with Timothy syndrome.

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We thank K. Timothy and the individuals with Timothy syndrome who participated in this study; E. Nigh for editing of the manuscript; U. Francke for karyotyping; A. Cherry and D. Bangs for help with fibroblast cultures; G. Panagiotakos and C. Young-Park for insightful discussions, and A. Krawisz, R. Schwemberger, D. Fu and R. Shu for help with data analysis. Antibodies to FORSE-1 were developed by P.H. Patterson and were obtained from the Developmental Studies Hybridoma Bank (University of Iowa). Financial support was provided by a US National Institutes of Health Director's Pioneer Award, and by grants to R.E.D. from the US National Institute of Mental Health, the California Institute for Regenerative Medicine and the Simons Foundation for Autism Research. S.P.P. was supported by awards from the International Brain Research Organization Outstanding Research Fellowship and the Tashia and John Morgridge Endowed Fellowship, M.Y. by a Japan Society of the Promotion for Science Postdoctoral Fellowship for Research Abroad and an American Heart Association Western States postdoctoral fellowship, T.P. by a Swiss National Science Foundation Postdoctoral Fellowship and A.S. by a California Institute for Regenerative Medicine Postdoctoral Fellowship. We are also grateful for funding from B. and F. Horowitz, M. McCafferey, B. and J. Packard, P. Kwan and K. Wang and the Flora foundation.

Author information

Author notes

    • Thomas Portmann
    • , Irina Voineagu
    •  & Masayuki Yazawa

    These authors contributed equally to this work.


  1. Department of Neurobiology, Stanford University School of Medicine, Stanford, California, USA.

    • Sergiu P Paşca
    • , Thomas Portmann
    • , Masayuki Yazawa
    • , Aleksandr Shcheglovitov
    • , Anca M Paşca
    •  & Ricardo E Dolmetsch
  2. Department of Neurology and Program in Neurogenetics, David Geffen School of Medicine, University of California, Los Angeles, California, USA.

    • Irina Voineagu
    •  & Daniel H Geschwind
  3. Stanford Institute for Stem Cell Biology and Regenerative Medicine and Department of Neurosurgery, Stanford School of Medicine, Stanford, California, USA.

    • Branden Cord
    •  & Theo D Palmer
  4. Department of Psychiatry and Behavioral Science, Stanford University School of Medicine, Stanford, California, USA.

    • Sachiko Chikahisa
    • , Seiji Nishino
    •  & Joachim Hallmayer
  5. Department of Integrative Physiology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, Japan.

    • Sachiko Chikahisa
  6. Department of Pediatrics, Stanford University School of Medicine, Stanford, California, USA.

    • Jonathan A Bernstein


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R.E.D. and S.P.P. designed the experiments and wrote the manuscript. S.P.P. generated iPSC lines, differentiated the iPSC lines into neurons, performed the calcium imaging and immunocytochemistry studies and contributed to the mutant mouse characterization. T.P. designed and analyzed the Fluidigm microarray studies. M.Y. generated and characterized the iPSC lines, and generated and characterized the mutant mice. I.V. and D.H.G. performed and analyzed the microarray gene expression experiments. A.S. derived neurons and designed and performed the electrophysiological experiments. A.M.P. performed the karyotyping and immunocytochemistry. S.C. and N.S. performed and analyzed catecholamine concentrations by HPLC. B.C. and T.D.P. contributed to the Fluidigm studies. J.A.B. and J.H. recruited and characterized the subjects.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Ricardo E Dolmetsch.

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    Supplementary Methods, Supplementary Figures 1–9 and Supplementary Tables 1–3 and 5

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    Genes differentially expressed between TS and controls

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