nmda receptor blockers  (Tocris)

Journal: Scientific Reports

Article Title: GluN2A-mediated currents and calcium signal in human iPSC-derived neurons

doi: 10.1038/s41598-026-38482-y

Figure Lengend Snippet: Electrophysiological measurements of NMDA-dependent currents in iPSC-derived neurons in BPM and NMM conditions after 60 days in culture. ( A ) AMPA and NMDA mediated components of synaptic currents were isolated by adding LY303070 , which selectively blocks AMPA-mediated EPSCs. Subsequent application of APV fully abolished spontaneous synaptic transmission. ( B ) Representative membrane current upon addition of 100 μM NMDA with 10 μM Glycine in neurons cultured in BPM and NMM. The superimposed grey line illustrates a single exponential fitting the desensitization process with the indicated τ. ( C , D ) Quantification of the NMDA-induced current amplitudes normalized respect to cell capacitance (C, n = 17 for BPM, n = 7 for NMM; p = 0.2094, Mann–Whitney test) and desensitization τ (D, n = 16 for BPM, n = 6 for NMM, p = 0.1775, Mann–Whitney test). ( E ) Same as B), upon addition of 1 μM Ro 25–6981 for inhibiting GluN2B containing receptors. Superimposed grey traces are the responses without blocker added. ( F ) Percentage of Ro 25–6981 inhibition respect to NMDA-induced current ( n = 15 for BPM, n = 5 for NMM, * p < 0.05, Two-tailed Student’s t test. Expression values are mean ± SEM).

Article Snippet: NMDA receptor blockers: APV (TOCRIS, 0106) 50 μM, and Ro 25-6981 1 μM, were added to NMDA 100 μM + Glycine 100 μM in free MgCl2 solution.

Techniques: Derivative Assay, Isolation, Transmission Assay, Membrane, Cell Culture, MANN-WHITNEY, Inhibition, Two Tailed Test, Expressing

Journal: Scientific Reports

Article Title: GluN2A-mediated currents and calcium signal in human iPSC-derived neurons

doi: 10.1038/s41598-026-38482-y

Figure Lengend Snippet: Intracellular calcium changes in iPSC-derived neural cultures in BPM and NMM conditions after 60 days in culture. ( A ) Representative average traces of the Fura-2 signal in neural cells cultured in BPM and NMM at basal conditions, after perfusion of the agonist NMDA (without Mg 2+ ), the addition of either the NMDAR antagonist APV ( n = 52 for BPM, n = 11 for NMM) or the GluN2B inhibitor Ro 25–6981 ( n = 33 for BPM and n = 11 for NMM), and finally perfusion of KCl. The pictures at the right side illustrate the Ca 2+ signal by Fura-2 at different time points indicated by coloured dots on the trace. ( B ) Quantification of Fura-2 signal (F340/380) under no stimulation in neural cultures ( n = 7 for BPM, n = 9 for NMM). ( C ) Percentage of cells that respond to NMDA ( n = 399 for BPM, n = 837 for NMM; deviation of > 0.1 in the Fura-2 signal from the baseline, **** p < 0.0001; Fisher’s exact test). ( D ) Amplitude of Fura-2 signal induced by NMDA perfusion ( n = 268 for BPM, n = 98 for NMM; **** p < 0.0001; Mann–Whitney test). ( E ) Percentage of Fura-2 signal inhibition by APV respect to NMDA Fura-2 signal ( n = 152 for BPM, n = 41 for NMM). ( F ) Percentage of Fura-2 signal inhibition induced by Ro-6981 respect to NMDA Fura-2 signal ( n = 117 for BPM, n = 58 for NMM; *** p < 0.0002; Mann–Whitney test). ( G ) Heatmap extracted from the RNA-seq showing the 45 up (red)- and down (blue)-regulated genes in the GO cluster “Calcium signaling”. Expression values are mean ± interquartile range.

Article Snippet: NMDA receptor blockers: APV (TOCRIS, 0106) 50 μM, and Ro 25-6981 1 μM, were added to NMDA 100 μM + Glycine 100 μM in free MgCl2 solution.

Techniques: Derivative Assay, Cell Culture, MANN-WHITNEY, Inhibition, RNA Sequencing, Expressing