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. 2016 Mar 22;11(3):e0151233. doi: 10.1371/journal.pone.0151233

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© 2016 Meunier et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 2 — Glycine is not the endogenous co-agonist of L5PyNs VC NMDARs A: Application of the glycine degrading enzyme BsGO (0.1 U/ml) has no effect on NMDA-EPSCs (n = 5 cells, 5 slices, 2 animals), indicating that glycine is not the endogenous co-agonist of synaptic L5PyrNs VC NMDARs. Scale bars: 100pA, 500ms. B: Further, enhancing endogenous glycine levels with the glycine transporter blocker ALX5407 (2 μM) decreased the NMDARs response, an effect blocked by BsGO (n = 4 cells, 4 slices, 2 animals). Scale bars: 25pA, 250ms. C: A similar result is obtained by exogenous application of glycine (100μM) (n = 5 cells, 5 slices, 2 animals). Scale bars: 50pA, 500ms. D: Such downregulation of NMDA-EPSCs by glycine is remarkably blocked by the glycinergic receptors (GyRs) antagonist strychnine (10μM) (n = 5 cells, 5 slices, 2 animals). Scale bars: 50pA, 500ms. E: Immunofluorescence for GlyRs revealed that, in the VC, they are mainly expressed in L5PyRNs notably at the somatic and dendritic level. Scale bar: 50μm, inset: 30μm. F: Altogether, these results indicate that glycine downregulates NMDA-EPSCs through activation of GlyRs ** p<0.01, *** p<0.001.