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Figure 4 | Neural Development

Figure 4

From: Semaphorin 5B mediates synapse elimination in hippocampal neurons

Figure 4

Overexpression of Sema5B reduces the number of synaptic inputs. (a-i) Confocal images of 13 days in vitro (DIV) hippocampal cultures cotransfected at 10 DIV with postsynaptic density (PSD)-95-RFP plus green fluorescent protein (GFP) (a), GFP-Sema5B (d), or GFP-Δsema (g), and immunolabeled for synaptophysin (b, e, h). Although the number of PSD-95-red fluorescent protein (RFP) puncta remained constant (c, f, i), there was a significant decrease in the number of synaptic inputs onto Sema5B overexpressing cells as measured by a decrease in the percentage of PSD-95 puncta that had an associated synaptophysin puncta (j; Student's t test; P < 0.005). Similarly there was a correlative decrease in the density of synaptic inputs (j; Student's t test; P < 0.05). The number and density of synaptic inputs onto cells overexpressing Sema5BΔsema was not significantly different than control (j; Student's t test; P > 0.5). The number of synapses being formed by Sema5B overexpressing cells, as measured by the number of PSD-95 puncta that colocalized with Syn-RFP (data not shown), was unchanged (k; Student's t test; P > 0.5). N = at least 15 neurons per condition from 3 separate cultures. Scale bar 20 μm.

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