NESH Regulates Dendritic Spine Morphology and Synapse Formation
Figure 4
Overexpression of NESH prevents synapse formation in hippocampal neurons.
(A) Cultured hippocampal neurons were co-transfected with GFP and myc-NESH at 10–12 DIV and fixed at 16–18 DIV. Empty vector was used as a control, and GFP was used to visualize dendritic spines. The fixed neurons were stained with anti-VAMP2 (presynaptic marker) antibody, anti-GluR1 (subunit of AMPA receptor) antibody or Alexa Fluor 594-conjugated phalloidin. (B) Synaptic densities were analyzed by counting the dendritic spines contacting presynapses marked by VAMP2 staining. (C) GluR1 clusters on dendritic spines were measured in neurons overexpressing NESH and compared with control. (D) F-actin fluorescence intensity ratios (spine vs. shaft). Data were obtained from three independent experiments; n = 20 each for control and NESH-overexpressing neurons. Data are presented as means ± SEM. *p<0.05, **p<0.01.