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Updated: Nov 13, 2025

A High-throughput Calcium-flux Assay to Study NMDA-receptors with Sensitivity to Glycine/D-serine and Glutamate
Published on: July 10, 2018
Protective effect of Achyranthes bidentata polypeptides on NMDA-mediated injury is developmentally regulated via
Wenqing Hu1,2, Jianghong He1, Yu Wang1
1Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, China.
Background:
Achyranthes bidentata polypeptides (ABPPs) are a potent intervention for excitotoxicity-related disorders such as Parkinson's disease and ischemic stroke. Previous work suggests that overstimulation of N-methyl-D-aspartate (NMDA) receptors plays a critical role in excitotoxicity, and expression of NR2 subunit variations is developmentally regulated. Our current study focused on neuroprotection of ABPPs on cultured neurons by modulation of NR2A and NR2B differentially.
Methods:
Primary cultured neurons were treated with NVP-AAM077, Ro-256981, ABPPs, and then the neurons were exposed to NMDA to induce excitotoxicity. Cellular viability was detected promptly and 24-hour after exposure to NMDA by MTT assay. Patch-clamp recording was applied to evaluate the effect of ABPPs on NMDA-evoked current and the differential modulation of ABPPs on NR2A and NR2B subunits in conjunction with NVP-AAM077 and Ro-256981.
Results:
ABPPs (10 µg/mL) blocked neuronal injury by NMDA in mature cultures, and the peptides conferred neuroprotection in immature cultures unless co-applied with NVP-AAM077. Furthermore, ABPPs enhanced NMDA current in mature cultures, while decreasing NMDA current in immature cultures. On the other hand, we showed that ABPPs increased NMDA current when Ro-256981 was present and decreased NMDA current when NVP-AAM007 was present.
Conclusions:
Neuroprotection of ABPPs on NMDA-mediated injury differentially in immature and mature cultures involves enhancement of NR2A subunits and prevention of NR2B subunits, indicating that dosage of ABPP should be considered in treatment with patients at different developmental stages.
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