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. 2012 Mar 21;32(12):3992–4003. doi: 10.1523/JNEUROSCI.4129-11.2012

Figure 5.

Figure 5.

Localization of GluN2B subunits. MSN (striatal neurons in corticostriatal coculture) and CTX (cortical neurons in corticostriatal coculture) were transfected with a YFP-tagged GluN2B subunit and grown in chambers for 21days in vitro. A, Representative examples of CTX (top) and MSNs (middle, bottom) transfected with a YFP-GluN2B subunit, live stained with α-GFP, fixed, and stained with α-PSD-95 as a postsynaptic marker and α-VGLUT1 as a presynaptic marker. B, Dendritic puncta density. CTX cells express more GluN2B-YFP clusters on secondary dendrites than MSNs. CTX and MSNs show similar distributions of VGLUT1 on secondary dendrites. MSNs show an increased density of PSD-95 on secondary dendrites compared with CTX. Significant by one-way ANOVA, F(5,120) = 18.46, p < 0.0001. Bonferroni's post-test, *p < 0.05,**p < 0.01. C, Dendritic puncta colocalization. The percentage colocalization between markers on dendrites was similar between CTX and MSN except for VGLUT1 colocalization with PSD-95, one-way ANOVA, F(5,120) = 8.00, p < 0.0001, Bonferroni's post-test, *p < 0.05. D, Somatic puncta density and colocalization. MSNs expressed more GluN2B on the soma than cortical cells. A greater percentage of VGLUT1 was associated with GluN2B in MSNs compared with cortical cells. One-way ANOVA, F(5,120) = 16.52, p < 0.0001, Bonferroni's post-test, *p < 0.05, **p < 0.01. B–D, n = 4–10 cells per culture from each of 3 separate cultures. E, Representative NMDAR currents from cocultured cortical neuron (left) or MSN (right), evoked by application of 1 mm NMDA (solid line) in control conditions, followed in the same neuron by NMDA application with treatment of 8 –12 min of 3 μm ifenprodil (Ifen). Scale bar, 1nA and 1s. F, Mean peak current densities in both cortical neurons (CTX; 38.00 ± 4.32, n = 8 from 3 cultures) and MSN (46.86 ± 4.25, n = 8 from 3 cultures) are reduced by ifenprodil (ifen) treatment (20.10 ± 2.87 in CTX, 23.97 ± 2.69 in MSNs, *p < 0.05 by one-way ANOVA, followed by Bonferroni's post-test).