Analysis of conditional heterozygous STXBP1 mutations in human neurons

Patzke, Christopher; Yan, Huaxiao; Covy, Jason P.; Yi, Fei; Maxeiner, Stephan; Wernig, Marius; Südhof, Thomas C.

doi:10.1172/jci78612

Cited by 73 publications

(83 citation statements)

References 44 publications

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“…They also used optogenetics (ChR2) and light stimulation of the enteric neurons to measure smooth muscle cell contractile responses [59]. A model for epileptic encephalopathy was made using gene-targeted ESCs, also in combination with optogenetics (ChR2/ChiEF) [60]. Together with the above-mentioned methods, modified iNGNs could also be created for disease model studies, along with ChR2 as a tool to precisely activate the iNGNs.…”

Section: Discussionmentioning

confidence: 99%

Functional Maturation of Human Stem Cell-Derived Neurons in Long-Term Cultures

et al. 2017

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Differentiated neurons can be rapidly acquired, within days, by inducing stem cells to express neurogenic transcription factors. We developed a protocol to maintain long-term cultures of human neurons, called iNGNs, which are obtained by inducing Neurogenin-1 and Neurogenin-2 expression in induced pluripotent stem cells. We followed the functional development of iNGNs over months and they showed many hallmark properties for neuronal maturation, including robust electrical and synaptic activity. Using iNGNs expressing a variant of channelrhodopsin-2, called CatCh, we could control iNGN activity with blue light stimulation. In combination with optogenetic tools, iNGNs offer opportunities for studies that require precise spatial and temporal resolution. iNGNs developed spontaneous network activity, and these networks had excitatory glutamatergic synapses, which we characterized with single-cell synaptic recordings. AMPA glutamatergic receptor activity was especially dominant in postsynaptic recordings, whereas NMDA glutamatergic receptor activity was absent from postsynaptic recordings but present in extrasynaptic recordings. Our results on long-term cultures of iNGNs could help in future studies elucidating mechanisms of human synaptogenesis and neurotransmission, along with the ability to scale-up the size of the cultures.

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Section: Discussionmentioning

confidence: 99%

Functional Maturation of Human Stem Cell-Derived Neurons in Long-Term Cultures

et al. 2017

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“…We co-expressed active (Cre) or mutant Cre-recombinase (ΔCre) with Ngn2 during neuronal differentiation to produce precisely matching wild-type (ΔCre [SHANK3 +/+ ]) and heterozygous mutant neurons (Cre [SHANK3 +/− ]; 24,25), using two independently targeted heterozygous clones of SHANK3 +/cKO ES cells to control for clonal variation (Fig. 1C).…”

Section: Generation Of Heterozygous Shank3 Conditional Knockout (Cko)mentioning

confidence: 99%

Autism-associated SHANK3 haploinsufficiency causes I _h channelopathy in human neurons

Dankó

Botelho

et al. 2016

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Heterozygous SHANK3 mutations are associated with idiopathic autism and Phelan-McDermid syndrome. SHANK3 is a ubiquitously expressed scaffolding protein that is enriched in postsynaptic excitatory synapses. Here we used engineered conditional mutations in human neurons to show that heterozygous and homozygous SHANK3 mutations severely and specifically impair Ih-channels. SHANK3 mutations caused alterations in neuronal morphology and synaptic connectivity; chronic pharmacological blockage of Ih-channels reproduced these phenotypes, suggesting they may be secondary to Ih-channel impairment. Moreover, mouse Shank3-deficient neurons also exhibited severe decreases in Ih-currents. SHANK3 protein interacted with hyperpolarization-activated cyclic nucleotide-gated channel proteins (HCN proteins) forming Ih-channels, indicating that SHANK3 functions to organize HCN-channels. Our data suggest SHANK3 mutations predispose to autism, at least partially, by inducing an Ih-channelopathy that may be amenable to pharmacological intervention.

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“…Heterozygous pathogenic variants in STXBP1 have been reported in individuals with epilepsy, intellectual disability, and autism . Human embryonic stem cell‐induced neurons with STXBP1 null mutations degenerate within 5 days, and heterozygous mutations result in a decrease of spontaneous and evoked excitatory neurotransmitter release, despite no change in soma size, dendritic morphology, or spine density . No animal model data have been reported yet.…”

Section: Neurexin and Neuroliginmentioning

confidence: 99%

Autism genetics – an overview

Yin

Schaaf

2016

Prenatal Diagnosis

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Autism spectrum disorder (ASD) is a highly heritable, clinically diverse group of neurodevelopmental disorders. Its genetic heterogeneity is remarkable, with more than 800 ASD predisposition genes identified to date. They are involved in various biological processes, including chromatin remodeling and gene transcription regulation, cell growth and proliferation, ubiquitination, and neuronal-specific processes, such as synaptic organization and activity, dendritic morphology, and axonogenesis. This review aims to discuss basic autism genetics, explicate ways to investigate ASD in model systems, highlight some key genes and their molecular pathways, and introduce novel theories of ASD pathogenesis, such as imbalance of excitatory and inhibitory brain activity, oligogenic heterozygosity, and the female protective model. © 2016 John Wiley & Sons, Ltd.

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Analysis of conditional heterozygous STXBP1 mutations in human neurons

Cited by 73 publications

References 44 publications

Functional Maturation of Human Stem Cell-Derived Neurons in Long-Term Cultures

Functional Maturation of Human Stem Cell-Derived Neurons in Long-Term Cultures

Autism-associated SHANK3 haploinsufficiency causes I _h channelopathy in human neurons

Autism genetics – an overview

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Analysis of conditional heterozygous STXBP1 mutations in human neurons

Cited by 73 publications

References 44 publications

Functional Maturation of Human Stem Cell-Derived Neurons in Long-Term Cultures

Functional Maturation of Human Stem Cell-Derived Neurons in Long-Term Cultures

Autism-associated SHANK3 haploinsufficiency causes I h channelopathy in human neurons

Autism genetics – an overview

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Product

Resources

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Autism-associated SHANK3 haploinsufficiency causes I _h channelopathy in human neurons