Journal Article

Rescuing tri-heteromeric NMDA receptor function: the potential of pregnenolone-sulfate in loss-of-function GRIN2B variants

Berlin, Shai, Kellner, Shai

2024

Overview

N-methyl-D-aspartate receptors (NMDARs emerging from GRIN genes) are tetrameric receptors that form diverse channel compositions in neurons, typically consisting of two GluN1 subunits combined with two GluN2(A-D) subunits. During prenatal stages, the predominant channels are di-heteromers with two GluN1 and two GluN2B subunits due to the high abundance of GluN2B subunits. Postnatally, the expression of GluN2A subunits increases, giving rise to additional subtypes, including GluN2A-containing di-heteromers and tri-heteromers with GluN1, GluN2A, and GluN2B subunits. The latter  emerge as the major receptor subtype at mature synapses in the hippocampus. Despite extensive research on purely di-heteromeric receptors containing two identical GRIN variants, the impact of a single variant on the function of other channel forms, notably tri-heteromers, is lagging. In this study, we systematically investigated the effects of two de novo GRIN2B variants (G689C and G689S) in pure, mixed di- and tri-heteromers. Our findings reveal that incorporating a single variant in mixed di-heteromers or tri-heteromers exerts a dominant negative effect on glutamate potency, although ‘mixed’ channels show improved potency compared to pure variant-containing di-heteromers. We show that a single variant within a receptor complex does not impair the response of all receptor subtypes to the positive allosteric modulator pregnenolone-sulfate (PS), whereas spermine completely fails to potentiate tri-heteromers containing GluN2A and -2B-subunits. We examined PS on primary cultured hippocampal neurons transfected with the variants, and observed a positive impact over current amplitudes and synaptic activity. Together, our study supports previous observations showing that mixed di-heteromers exhibit improved glutamate potency and extend these findings towards the exploration of the effect of Loss-of-Function variants over tri-heteromers. Notably, we provide an initial and crucial demonstration of the beneficial effects of GRIN2B -relevant potentiators on tri-heteromers. Our results underscore the significance of studying how different variants affect distinct receptor subtypes, as these effects cannot be inferred solely from observations made on pure di-heteromers. Overall, this study contributes to ongoing efforts to understand the pathophysiology of GRINopathies and provides insights into potential treatment strategies.

• Publisher: Springer International Publishing,Springer Nature B.V
• Subject: Allosteric properties / Animals / Biochemistry / Biomedical and Life Sciences / Biomedicine / Brain / Cell Biology / Channels / Glutamate receptors / glutamic acid / Glutamic acid receptors (ionotropic) / HEK293 Cells / Hippocampus / Hippocampus - metabolism / Humans / Life Sciences / Loss of Function Mutation / N-Methyl-D-aspartic acid receptors / Neurons / Neurons - metabolism / Original / Original Article / pathophysiology / Pregnenolone / Pregnenolone - metabolism / Pregnenolone - pharmacology / Protein Multimerization / Protein Subunits - genetics / Protein Subunits - metabolism / Receptor mechanisms / Receptors / Receptors, N-Methyl-D-Aspartate - genetics / Receptors, N-Methyl-D-Aspartate - metabolism / Spermine / Sulfates / Synapses