A NMDA-receptor calcium influx assay sensitive to stimulation by glutamate and glycine/D-serine.
A NMDA-receptor calcium influx assay sensitive to stimulation by glutamate and glycine/D-serine.
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DOI：
10.1038/s41598-017-11947-x
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发表时间：
2017-09-14
影响因子：
4.6
通讯作者：
Bill A
中科院分区：
文献类型：
--
作者：
Guo H;Camargo LM;Yeboah F;Digan ME;Niu H;Pan Y;Reiling S;Soler-Llavina G;Weihofen WA;Wang HR;Shanker YG;Stams T;Bill A
N-甲基-D-天冬氨酸受体（NMDAR）是离子型谷氨酸受体，其在突触传递、可塑性和认知中起作用。NMDAR的功能障碍与多种神经系统疾病有关，使其成为有吸引力的治疗靶点。功能性NMDAR在非神经元细胞中的过表达通过兴奋性毒性导致细胞死亡，阻碍了用于NMDAR药物发现的基于细胞的测定的发展。在这里，我们报告了一个基于板的，高通量的方法来研究NMDAR功能。我们的测定使得能够在甘氨酸/D-丝氨酸或谷氨酸激活后对具有不同亚基组成的NMDAR进行功能研究，因此提出了第一个基于板的高通量测定，其允许在甘氨酸/D-丝氨酸和/或谷氨酸敏感模式下测量NMDAR功能。这允许研究小分子调节剂在不同浓度或共配体组合下对NMDAR活化的影响。所报道的测定系统忠实地复制了受体对已知激动剂、拮抗剂、正变构调节剂和负变构调节剂的药理学反应，以及受体对镁和锌的敏感性。我们相信，快速和大规模筛选研究NMDAR生物学的能力将能够鉴定新的治疗剂，其发现否则会受到现有基于细胞的方法的局限性的阻碍。
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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影响因子：
64.8
作者：
Gielen, Marc;Retchless, Beth Siegler;Mony, Laetitia;Johnson, Jon W.;Paoletti, Pierre
通讯作者：
Paoletti, Pierre
影响因子：
5
作者：
Feuerbach, Dominik;Loetscher, Erika;Koller, Manuel
通讯作者：
Koller, Manuel
影响因子：
3.3
作者：
Horak, M;Vlcek, K;Vyklicky, L
通讯作者：
Vyklicky, L
影响因子：
7.3
作者：
Feng, BH;Tse, HW;Monaghan, DT
通讯作者：
Monaghan, DT
DOI：
10.1073/pnas.1217072110
发表时间：
2013-03-12
影响因子：
11.1
作者：
Britschgi, Adrian;Bill, Anke;Bentires-Alj, Mohamed
通讯作者：
Bentires-Alj, Mohamed