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Fig. 5. ApSyn induces ultrastructural changes in C1 neurons in a phosphorylation-dependent manner. (A-C) Fine structure of contact areas between C1 and C3 neurons of soma-soma pairs in which C1 has been injected with GST (A-A"), apSyn (B-B") or apSynALA9 (C-C"). The boundaries of the meshwork of interdigitating cellular processes between C1 and C3 somata are highlighted in black (arrows). Note the marked increase in the thickness of this area in the sample injected with apSyn (B-B') with respect to either GST- or apSynALA9-loaded samples (A-A' and C-C', respectively). In the apSyn-injected samples the area of contact was characterized by a dense meshwork of neurite-like processes filled with microtubules and clusters of dense core SV profiles typical of the C1 neuron (see arrowhead in B and high magnification inset). Clusters of SVs were also visible in processes extending outside the contact area (B"). These processes were virtually absent in C1-C3 pairs loaded with GST or apSynALA9 (A-A" and C-C"). (D) Bar graph showing the differences in the thickness of the meshwork of embricated processes between C1 and C3 in the three experimental groups. When the C1 neuron was loaded with apSyn, the interdigitation area was significantly thicker than in control conditions (GST injection) or after injection of the non-phosphorylatable mutant apSynALA9. (E) A cluster of dense core SVs typical of the C1 neuron. (F) Large neurosecretory granules observed in the soma of C3. Bar, 1.8 µm for panels A-C; 8 µm for panels A'-C'; 1 µm for panels A"-C"; 0.5 µm for panels E,F and inset in panel B.
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