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Stem Cell Reports. 2018 Jul 10;11(1):183-196. doi: 10.1016/j.stemcr.2018.05.018. Epub 2018 Jun 21.

Disruption of GRIN2B Impairs Differentiation in Human Neurons.

Author information

1
McGill University and Douglas Hospital Research Institute, Department of Psychiatry, 6875 LaSalle Boulevard, Frank Common Building, Room 2101.2, Verdun, Montreal, QC H4H 1R3, Canada.
2
Montreal Neurological Institute, Department of Neurology and Neurosurgery, Montreal, QC H3A 2B4, Canada.
3
Department of Pediatrics, Developmental Biology Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Northwestern University, Feinberg School of Medicine, Chicago, IL 60611, USA.
4
McGill University and Douglas Hospital Research Institute, Department of Psychiatry, 6875 LaSalle Boulevard, Frank Common Building, Room 2101.2, Verdun, Montreal, QC H4H 1R3, Canada. Electronic address: carl.ernst@mcgill.ca.

Abstract

Heterozygous loss-of-function mutations in GRIN2B, a subunit of the NMDA receptor, cause intellectual disability and language impairment. We developed clonal models of GRIN2B deletion and loss-of-function mutations in a region coding for the glutamate binding domain in human cells and generated neurons from a patient harboring a missense mutation in the same domain. Transcriptome analysis revealed extensive increases in genes associated with cell proliferation and decreases in genes associated with neuron differentiation, a result supported by extensive protein analyses. Using electrophysiology and calcium imaging, we demonstrate that NMDA receptors are present on neural progenitor cells and that human mutations in GRIN2B can impair calcium influx and membrane depolarization even in a presumed undifferentiated cell state, highlighting an important role for non-synaptic NMDA receptors. It may be this function, in part, which underlies the neurological disease observed in patients with GRIN2B mutations.

KEYWORDS:

CRISPR; CRISPR-Cas9; GRIN2B; NMDA; NMDAR2B; NPCs; glutamate; iPSCs; neural stem cell; neurodevelopment

PMID:
29937144
PMCID:
PMC6067152
DOI:
10.1016/j.stemcr.2018.05.018
[Indexed for MEDLINE]
Free PMC Article

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