Abstract
OBJECTIVE: GRIN1 -related neurodevelopmental disorder ( GRIN1 -NDD) is characterized by clinically significant variation in the GRIN1 gene, which encodes the obligatory GluN1 subunit of N-methyl-D-aspartate receptors (NMDARs). The identified p.Tyr647Ser (Y647S) variant - carried by a 33-year-old female with seizures and intellectual disability - is located in the M3 helix in the GluN1 transmembrane domain. This study builds upon initial in vitro investigations of the functional impacts of the GRIN1 Y647S variant and examines its in vivo consequences in a mouse model. METHODS: To investigate in vitro functional impacts of NMDARs containing GluN1-Y647S variant subunits, GluN1-Y647S was co-expressed with wildtype GluN2A or GluN2B subunits in Xenopus laevis oocytes and HEK cells. Grin1 (Y647S/+) mice were created by CRISPR-Cas9 endonuclease-mediated transgenesis and the molecular, electrophysiological, and behavioural consequences of the variant were examined. RESULTS: In vitro , NMDARs containing GluN1-Y647S show altered sensitivity to endogenous agonists and negative allosteric modulators, and reduced cell surface trafficking. Grin1 (Y647S/+) mice displayed a reduction in whole brain GluN1 levels and deficiency in NMDAR-mediated synaptic transmission in the hippocampus. Behaviourally, Grin1 (Y647S/+) mice exhibited spontaneous seizures, altered vocalizations, muscle strength, sociability, and problem-solving. INTERPRETATION: The Y647S variant confers a complex in vivo phenotype, which reflects largely diminished properties of NMDAR function. As a result, Grin1 (Y647S/+) mice display atypical behaviour in domains relevant to the clinical characteristics of GRIN1 -NDD and the individual carrying the variant. Ultimately, the characterization of Grin1 (Y647S/+) mice accomplished in the present work expands our understanding of the mechanisms underlying GRIN1 -NDD and provides a foundation for the development of novel therapeutics.