A NMDA-receptor Calcium Influx Assay Sensitive to Stimulation by Glutamate and Glycine/D-serine

Overview

Journal Sci Rep

Specialty Science

Date 2017 Sep 16

PMID 28912557

Citations 22

Authors

Hongqiu Guo

L Miguel Camargo

Fred Yeboah

Mary Ellen Digan

Honglin Niu

Yue Pan

Stephan Reiling

Gilberto Soler-Llavina

Wilhelm A Weihofen

Hao-Ran Wang

Y Gopi Shanker

Travis Stams

Anke Bill

Affiliations

Soon will be listed here.

Abstract

N-methyl-D-aspartate-receptors (NMDARs) are ionotropic glutamate receptors that function in synaptic transmission, plasticity and cognition. Malfunction of NMDARs has been implicated in a variety of nervous system disorders, making them attractive therapeutic targets. Overexpression of functional NMDAR in non-neuronal cells results in cell death by excitotoxicity, hindering the development of cell-based assays for NMDAR drug discovery. Here we report a plate-based, high-throughput approach to study NMDAR function. Our assay enables the functional study of NMDARs with different subunit composition after activation by glycine/D-serine or glutamate and hence presents the first plate-based, high throughput assay that allows for the measurement of NMDAR function in glycine/D-serine and/or glutamate sensitive modes. This allows to investigate the effect of small molecule modulators on the activation of NMDARs at different concentrations or combinations of the co-ligands. The reported assay system faithfully replicates the pharmacology of the receptor in response to known agonists, antagonists, positive and negative allosteric modulators, as well as the receptor's sensitivity to magnesium and zinc. We believe that the ability to study the biology of NMDARs rapidly and in large scale screens will enable the identification of novel therapeutics whose discovery has otherwise been hindered by the limitations of existing cell based approaches.

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