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Fig. 3. Synaptogenesis is impaired in Dyn3baa-expressing neurons. Neurons transfected with Dyn3aaa-GFP (A,C) or Dyn3baa-GFP (B,D) at 18DIV were labeled for the presynaptic marker synapsin (A,B) or the postsynaptic marker PSD-95 (C,D). Dyn3aaa-GFP was found to be directly juxtaposed to synapsin-positive puncta (arrows in A) and colocalized with PSD-95 (arrows in C). Dyn3baa-GFP and dendritic filopodia were neither directly juxtaposed to synapsin-positive puncta (arrows in B) nor colocalized with PSD-95 (arrows in D). (E,F) Quantitation of association between dendritic protrusions and synaptic markers revealed that control neurons exhibited little association with synaptic markers at 11DIV but extensive association at 18DIV. By contrast, neurons transfected with Dyn3baa-GFP exhibited impaired dendritic spine synaptogenesis, with 20% of filopodia containing PSD-95 (E) and less than 10% of filopodia juxtaposed to synapsin (F) at 18DIV. Neurons expressing a GTPase-deficient mutant Dyn3baa-GFP (KA mutant) did not exhibit this maturation defect and progressed normally through synaptogenesis with culture age. All error bars represent s.d. Scale bar, 5 µm.





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