Journal: Scientific Reports

Article Title: Stress hormone rapidly tunes synaptic NMDA receptor through membrane dynamics and mineralocorticoid signalling

doi: 10.1038/s41598-017-08695-3

Figure Lengend Snippet: Corticosterone alters surface GluN2B-NMDAR clustering. ( A ) Dendritic fragments of GluN2A- and GluN2B-SEP expressing neurons before and after exposure to corticosterone (100 nM). Scale bar = 5 µm, scale bar inset = 1 µm. ( B ) Example of fluorescence intensity of GluN2A- (n = 12 dendritic fields, N = 5 neurons) and GluN2B-SEP (n = 15 dendritic fields, N = 5 neurons) clusters over time. ( C ) Comparison of GluN2A- and GluN2B-SEP cluster fluorescence intensity before and after exposure to corticosterone. ***p < 0.001, paired Student t-test. ( D ) Live immunostaining of GluN2A and GluN2B subunits in Homer 1c-DsRed expressing neurons. Scale bar = 10 µm, scale bar inset = 5 µm. ( E ) Immunostaining of surface GluN2A and GluN2B subunits before and after exposure to corticosterone. Scale bar = 5 µm. ( F ) Comparison of the fluorescence intensity of GluN2A (n = 12 dendritic fields, N = 5 neurons) and GluN2B (n = 11 dendritic fields, N = 5 neurons) subunit clusters (expressed as ratio) -SEP before and after exposure to corticosterone (50 nM, 20 min). *p < 0.05, Student t-test.

Article Snippet: In a separate series of experiments, testing the involvement of GluN2B subunits, the selective GluN2B- blocker Ro 25–6981 (3 μM, Tocris) or vehicle (DMSO) was also directly added into the extracellular solution while recording.

Techniques: Expressing, Fluorescence, Comparison, Immunostaining

Journal: Scientific Reports

Article Title: Stress hormone rapidly tunes synaptic NMDA receptor through membrane dynamics and mineralocorticoid signalling

doi: 10.1038/s41598-017-08695-3

Figure Lengend Snippet: Corticosterone specifically decreases the surface dynamics of GluN2B-NMDAR in hippocampal neurons exposed to corticosterone. ( A ) Representative trajectories of single GluN2A- and GluN2B-NMDAR in hippocampal neurons (DIC images) exposed to either buffer or corticosterone (100 nM, 20 min). Scale bar, 5 µm; scale bar inset, 1 µm. ( B ) Comparison of the cumulative distributions of GluN2A- (buffer, n = 112 trajectories; corticosterone, n = 143 trajectories) and GluN2B-NMDAR (buffer, n = 189 trajectories; corticosterone, n = 191 trajectories) instantaneous diffusion coefficients in buffer and corticosterone conditions. ***p < 0.001, Kolmogorov-Smirnov test.

Article Snippet: In a separate series of experiments, testing the involvement of GluN2B subunits, the selective GluN2B- blocker Ro 25–6981 (3 μM, Tocris) or vehicle (DMSO) was also directly added into the extracellular solution while recording.

Techniques: Comparison, Diffusion-based Assay

Journal: Scientific Reports

Article Title: Stress hormone rapidly tunes synaptic NMDA receptor through membrane dynamics and mineralocorticoid signalling

doi: 10.1038/s41598-017-08695-3

Figure Lengend Snippet: Aldosterone, a MR activator, alters the surface distribution and dynamics of GluN2B-NMDAR. ( A ) Dendritic fragments of GluN2B-SEP expressing neurons before and after exposure to aldosterone (10 nM). Scale bar = 5 µm, scale bar inset = 1 µm. ( B ) Example of fluorescence intensity of GluN2B-SEP clusters over time, before and after aldosterone application. ( C ) Comparison of GluN2B-SEP cluster fluorescence intensity before and after exposure to aldosterone (10 nM, 25 min exposure; n = 10 dendritic fields, N = 5 neurons). ***p < 0.001, paired Student t-test. ( D ) Representative trajectories of single GluN2B-NMDAR in hippocampal neurons (DIC images) exposed to either buffer solution, aldosterone (10 nM, 20 min), corticosterone BSA (50 nM, 20 min), or RU28392 (GR agonist, 50 nM, 20 min). Scale bar, 5 µm. ( E ) Comparison of the cumulative distributions of GluN2B-NMDAR instantaneous diffusion coefficients between aldosterone (buffer, n = 214 trajectories; aldosterone, n = 225 trajectories), corticosterone BSA (buffer, n = 539 trajectories; 5min corticosterone BSA, n = 855 trajectories; 20min corticosterone BSA, n = 864 trajectories), or RU28392 (buffer, n = 229 trajectories; 5min RU28392, n = 175 trajectories; 20min RU28392, n = 119 trajectories). ***p < 0.001, Kolmogorov-Smirnov test.

Article Snippet: In a separate series of experiments, testing the involvement of GluN2B subunits, the selective GluN2B- blocker Ro 25–6981 (3 μM, Tocris) or vehicle (DMSO) was also directly added into the extracellular solution while recording.

Techniques: Expressing, Fluorescence, Comparison, Diffusion-based Assay

Journal: Scientific Reports

Article Title: Stress hormone rapidly tunes synaptic NMDA receptor through membrane dynamics and mineralocorticoid signalling

doi: 10.1038/s41598-017-08695-3

Figure Lengend Snippet: Corticosterone and aldosterone decrease GluN2B-NMDAR surface dynamics within synapses. ( A ) Representative trajectories of surface GluN2B-NMDAR in hippocampal synapses exposed to either buffer solution, aldosterone (10 nM, 20 min), or corticosterone (100 nM, 20 min). Scale bar, 200 nm. ( B ) Comparison of the synaptic GluN2B-NMDAR instantaneous diffusion coefficients between buffer and corticosterone (buffer, n = 141 trajectories; corticosterone, n = 138 trajectories), and buffer and aldosterone (buffer, n = 291 trajectories; aldosterone, n = 402 trajectories). Data are expressed as median diffusion coefficient ±25–75% IQR. *p < 0.05, two-tailed Mann-Whitney test.

Article Snippet: In a separate series of experiments, testing the involvement of GluN2B subunits, the selective GluN2B- blocker Ro 25–6981 (3 μM, Tocris) or vehicle (DMSO) was also directly added into the extracellular solution while recording.

Techniques: Comparison, Diffusion-based Assay, Two Tailed Test, MANN-WHITNEY