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Figure 4 | Neural Development

Figure 4

From: Neuroligin1: a cell adhesion molecule that recruits PSD-95 and NMDA receptors by distinct mechanisms during synaptogenesis

Figure 4

Patching endogenous Nlg induced clustering of PSD-95. (A) Patching with β-NRX-Fc (patched) increased the density of PSD-95 clusters as shown by confocal images of neurons that were incubated with β-NRX-Fc (red) for 1 hour then fixed and immunostained for PSD-95 (green). Secondary antibody only was used as the control. Scale bar, 20 μm. (B, C) Enlargements of the boxed regions in (A), showing control and patched dendrites, respectively. Scale bar, 10 μm. (D) Time-course of PSD-95 clustering by β-NRX-Fc demonstrates that the increase in PSD-95 cluster density occurred after 30 minutes of patching. (E) The density of PSD-95 clusters (per 20 μm of dendrite) was increased by patching Nlg, but not in the presence of 2-bromopalmitate (*P < 0.004). (F) Live imaging of a cortical neuron during patching with β-NRX-Fc demonstrates the appearance of new PSD-95-GFP clusters (yellow arrows). Sites at which clusters appeared are denoted with ghost arrows in the first image. Time is in hours and minutes. Scale bar, 5 μm. (G) Time-course of fluorescence intensities from five neurons treated with β-NRX-Fc. The locations at which intensities were measured have been categorized into sites with stable fluorescence (constant), sites at which clusters appear (appear) and sites at which clusters disappear (disappear). (H) Analysis of the numbers of clusters per neuron as categorized in (G). Error bars are standard error.

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