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• W3092532983 abstract "Replication studies play an essential role in corroborating research findings and ensuring that subsequent experimental works are interpreted correctly. A previously published paper indicated that the neurotransmitter glutamate, along with the compounds <i>N</i>-methyl-d-aspartate (NMDA) and d-(−)-2-amino-5-phosphonopentanoic acid (AP5), acts as positive allosteric modulators of inhibitory glycine receptors. The paper further suggested that this form of modulation would play a role in setting the spinal inhibitory tone and influencing sensory signaling, as spillover of glutamate onto nearby glycinergic synapses would permit rapid crosstalk between excitatory and inhibitory synapses. Here, we attempted to replicate this finding in primary cultured spinal cord neurons, spinal cord slice, and <i>Xenopus laevis</i> oocytes expressing recombinant human glycine receptors. Despite extensive efforts, we were unable to reproduce the finding that glutamate, AP5, and NMDA positively modulate glycine receptor currents. We paid careful attention to critical aspects of the original study design and took into account receptor saturation and protocol deviations such as animal species. Finally, we explored possible explanations for the experimental discrepancy. We found that solution contamination with a high-affinity modulator such as zinc is most likely to account for the error, and we suggest methods for preventing this kind of misinterpretation in future studies aimed at characterizing high-affinity modulators of the glycine receptor. <h3>SIGNIFICANCE STATEMENT</h3> A previous study indicates that glutamate spillover onto inhibitory synapses can directly interact with glycine receptors to enhance inhibitory signalling. This finding has important implications for baseline spinal transmission and may play a role when chronic pain develops. However, we failed to replicate the results and did not observe glutamate, d-(−)-2-amino-5-phosphonopentanoic acid, or <i>N</i>-methyl-d-aspartate modulation of native or recombinant glycine receptors. We ruled out various sources for the discrepancy and found that the most likely cause is solution contamination." @default.
• W3092532983 created "2020-10-15" @default.
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• W3092532983 date "2020-10-13" @default.
• W3092532983 modified "2023-10-16" @default.
• W3092532983 title "Glutamate, d-(−)-2-Amino-5-Phosphonopentanoic Acid, and N-Methyl-d-Aspartate Do Not Directly Modulate Glycine Receptors" @default.
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• W3092532983 doi "https://doi.org/10.1124/molpharm.120.000127" @default.
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• W3092532983 hasPublicationYear "2020" @default.
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