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Figure 4 | Neural Development

Figure 4

From: Role of the Wnt receptor Frizzled-1 in presynaptic differentiation and function

Figure 4

Fz1 overexpression induces presynaptic protein clustering. (A) The number of Bassoon clusters per neurite length was evaluated in hippocampal neurons (9 DIV) treated for 24 h with different concentrations of the soluble extracellular Fz1 CRD or 1,000 ng/ml Fz2-CRD. (B) Representative images of control neurons and neurons treated with 1 μg/ml Fz1-CRD or Fz2-CRD for 24 h. (C) Hippocampal neurons were transfected with Fz1-GFP, which was detected in axons stained with phosphorylated MAP1B (MAP1BP). (D) Fz1-GFP clusters co-localize with pre-synaptic terminals loaded with FM4-64 under stimulation with 90 mM KCl (arrows). (E) Hippocampal neurons were transfected with Fz1-myc plus GFP to easily identify transfected neurons. Fz1 staining (red) is highly increased in transfected neurons (asterisk) compared to non-transfected neurons (arrow). (F) Bassoon puncta were analyzed in neurons transfected with Fz1-myc or with empty vector (pCS2+) using an anti-Bassoon antibody and quantifying the clusters per neurite length in GFP-positive axons. There was an increased number of Bassoon puncta in neurons transfected with Fz1-myc compared to control. *P < 0.05; **P < 0.01; NS, not significant. Error bars indicate standard error of the mean.

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