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A high-throughput calcium-flux assay to study NMDAR-receptors with sensitivity to Glycine/D-Serine an Glutamate

Yeboah, Fred and Guo, Hongqiu and Bill, Anke (2018) A high-throughput calcium-flux assay to study NMDAR-receptors with sensitivity to Glycine/D-Serine an Glutamate. Journal of visualized experiments : JoVE, 137 (e58160). ISSN 1940-087X

Abstract

N-methyl-D-aspartate (NMDA) receptors are ionotropic glutamate receptors that function in learning and memory. NMDAR malfunction, expressed either as over activity or lack of activity caused by mutations, altered expression, trafficking or localization can contribute to numerous diseases especially in the central nervous system. Therefore, understanding the biology of the receptor as well as facilitating the discovery of compounds or small molecules is of exquisite importance in ongoing efforts to combat neurological diseases. Current approaches to study the receptor have limitations such as low throughput, high cost and the inability to study the functional abilities of the receptor due to the necessary presence of channel blockers to prevent NMDAR-mediated excitotoxicity. Additionally, the existing assay systems have a limitation of being sensitive to stimulation by glutamate only, lacking sensitivity to stimulation with glycine, one of the two co-ligands of the NMDA receptor.

Here we present the first plate-based high throughput approach to study NMDA receptor with sensitivity to both co-ligands; glutamate and D-serine/glycine. This approach allows the study of different compositions of NMDAR subunits and allows functional studies of the receptor in glycine and or glutamate sensitive modes and does not require the presence of inhibitors during the measurement. The effect of positive and negative allosteric modulators can be detected with this assay and the known pharmacology of NMDAR has been replicated in our system. This technique overcomes the limitations of existing methods, is cost effective and facile. We believe this novel technique will speed up the discovery of therapies for NMDAR-mediated pathologies.

Item Type: Article
Date Deposited: 07 Aug 2018 00:45
Last Modified: 07 Aug 2018 00:45
URI: https://oak.novartis.com/id/eprint/35708

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