Fig. 4.

Cooperation of N-cadherin and neuroligin-1 in CA3 pyramidal neurons. (A) Three-dimensional reconstruction of an EGFP expressing CA3 pyramidal neuron in a rat hippocampal slice culture (transfected by single cell electroporation). (Scale bar, 10 μm.) (B–D) Neuroligin-1-EGFP (Nlg-GFP) or EGFP were coelectroporated with dominant-negative N-cadΔE (ectodomain deleted) into single cells at 5 DIV and presynaptic vesicle clusters were immunostained for VAMP2 2 d later at 7 DIV. (B) Three-dimensional images of proximal dendrites. VAMP2 puncta on EGFP expressing dendrites are indicated by arrows. (Scale bar, 3 μm.) (C) Density of VAMP2 puncta on dendrites. (D) Change in VAMP2 puncta density upon neuroligin-1-EGFP expression depends on N-cadherin function. (E and F) Neuroligin-1-EGFP clustering depends on N-cadherin function. (E) (Upper) Original images of neuroligin-1-EGFP-expressing dendrites. (Lower) Thresholded neuroligin-1-EGFP puncta. (Scale bars: 2 μm.) (F) Dendritic density of neuroligin-1-EGFP puncta. Mean ± SEM; *P < 0.05; **P < 0.01, unpaired t test.