Wide phenotypic spectrum of human stem cell-derived excitatory neurons with Rett syndrome-associated MECP2 mutations
Wide phenotypic spectrum of human stem cell-derived excitatory neurons with Rett syndrome-associated MECP2 mutations
ABSTRACT Rett syndrome (RTT) is a severe neurodevelopmental disorder primarily caused by heterozygous loss-of-function mutations in the X-linked gene methyl-CpG-binding protein 2 (MECP2) that is a global transcriptional regulator. Mutations in the methyl-binding domain (MBD) of MECP2 disrupt its interaction with methylated DNA required for proper function in the brain. Here, we investigate the effect of a novel MECP2 L124W missense mutation in the MBD in comparison to MECP2 null mutations. L124W protein had a limited ability to disrupt heterochromatic chromocenters due to decreased binding dynamics. We isolated two pairs of isogenic WT and L124W induced pluripotent stem cell lines. L124W induced excitatory neurons expressed stable protein, exhibited only increased input resistance and impaired voltage-gated Na+ and K+ currents, and their neuronal dysmorphology was limited to reduced dendritic complexity. Three isogenic pairs of MECP2 null neurons had the expected more pronounced morphological and electrophysiological phenotypes, exhibiting decreased soma area, dendrite length, capacitance and excitatory synaptic function. We examined development and maturation of excitatory neural networks using micro-electrode arrays to detect alterations in RTT connectivity. The L124W neurons had no detectable changes in network circuitry features, in contrast to MECP2 null neurons that suffered a significant change in synchronous network burst frequency and a transient extension of network burst duration. Our results from stem cell-derived RTT excitatory neurons reveal a wide range of morphological, electrophysiological and circuitry phenotypes that reflect the severity of the MECP2 mutation.
Salter Michael W、Mok Rebecca SF、Zhang Wenbo、Rodrigues Deivid C、Wei Wei、Vincent John B、Muotri Alysson R、Sheikh Taimoor I、Mufteev Marat、Piekna Alina、Liu Jiaje、Ellis James、Fernandes Isabella R、Hildebrandt Matthew R、DeJong Leah C
Neurosciences & Mental Health Program, The Hospital for Sick ChildrenDepartment of Molecular Genetics, University of Toronto||Developmental & Stem Cell Biology Program, The Hospital for Sick ChildrenNeurosciences & Mental Health Program, The Hospital for Sick ChildrenDevelopmental & Stem Cell Biology Program, The Hospital for Sick ChildrenDevelopmental & Stem Cell Biology Program, The Hospital for Sick ChildrenMolecular Neuropsychiatry & Development (MiND) Lab, Campbell Family Mental Health Research Institute, Centre for Addiction and Mental HealthUniversity of California San Diego, School of Medicine, Department of Pediatrics/Rady Children?ˉs Hospital San Diego, Department of Cellular & Molecular Medicine, Stem Cell ProgramMolecular Neuropsychiatry & Development (MiND) Lab, Campbell Family Mental Health Research Institute, Centre for Addiction and Mental HealthDepartment of Molecular Genetics, University of Toronto||Developmental & Stem Cell Biology Program, The Hospital for Sick ChildrenDevelopmental & Stem Cell Biology Program, The Hospital for Sick ChildrenDevelopmental & Stem Cell Biology Program, The Hospital for Sick ChildrenDepartment of Molecular Genetics, University of Toronto||Developmental & Stem Cell Biology Program, The Hospital for Sick ChildrenUniversity of California San Diego, School of Medicine, Department of Pediatrics/Rady Children?ˉs Hospital San Diego, Department of Cellular & Molecular Medicine, Stem Cell ProgramDevelopmental & Stem Cell Biology Program, The Hospital for Sick ChildrenDepartment of Molecular Genetics, University of Toronto||Developmental & Stem Cell Biology Program, The Hospital for Sick Children
神经病学、精神病学基础医学分子生物学
Salter Michael W,Mok Rebecca SF,Zhang Wenbo,Rodrigues Deivid C,Wei Wei,Vincent John B,Muotri Alysson R,Sheikh Taimoor I,Mufteev Marat,Piekna Alina,Liu Jiaje,Ellis James,Fernandes Isabella R,Hildebrandt Matthew R,DeJong Leah C.Wide phenotypic spectrum of human stem cell-derived excitatory neurons with Rett syndrome-associated MECP2 mutations[EB/OL].(2025-03-28)[2025-04-26].https://www.biorxiv.org/content/10.1101/2020.07.12.189621.点此复制
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