Skip to main content
. Author manuscript; available in PMC: 2017 Jul 1.
Published in final edited form as: Neurobiol Dis. 2016 Mar 5;91:166–181. doi: 10.1016/j.nbd.2016.03.003

Figure 2.

Figure 2

Putative excitatory synapses on Pyr cell somata. A: Confocal image stack reconstruction (0.25 μm optical distance) of biocytin-filled control layer V pyramidal neuron (green). Section processed with antibodies for VGLUT1 and PSD95 such that dual co-immunoreactivity, selected by Volocity software, is magenta. White arrowheads point to sites of closely apposed VGLUT1- and PSD95-IR along basilar dendrites. B: Biocytin-filled layer V pyramidal cell from cortex 3d after undercut, reconstructed as in cell of A. Voxels of VGLUT1/PSD95 (magenta) in close apposition on basal dendrites (white arrowheads) and soma (open arrow) are indicated and were confirmed during cell rotation (Methods and Fig. S1). In A and B, only VGLUT1/PSD95-IR in close apposition to green cell is labeled magenta. C: A biocytin-labeled layer V pyramidal cell (green) from 3d undercut cortex. Two adjacent images (optical distance 0.5 μm) were obtained through the center of the cell body and collapsed. VGLUT1- (red) and PSD95-IR (blue) are also shown. Black void (lower right) corresponds to soma of adjacent unfilled cell. Lower white rectangle: proximal apical dendrite. D: Enlarged image of portion of soma and basal dendrite within the upper white rectangle of C. Black arrows point to sites of close apposition of VGLUT1/PSD95-IR on soma (arrows 1–3) and dendrite (arrow 4). D1, D2: Enlarged areas of close apposition (presumptive synapses) indicated by arrows 1 and 2 in D, respectively. Calibration: in A and B: 50 μm; in C: 10 μm. Green: biocytin; red: VGLUT1; blue: PSD95. E: 1.0 μm optical section cut through the soma of a layer V pyramidal cell from a P21, 3 day UC rat perfused with 4% paraformaldehyde and reacted for PSD95 (green), VGLUT1 (red) and NeuN (blue soma). White arrows: colocalization (yellow spots) of post-synaptic PSD95 and presynaptic VGLUT1 on the soma at presumptive excitatory synapses. Yellow arrowhead upper right: VGLUT1 and PSD95 colocalization in neuropil. E1: Enlargement of area in white box of E ~12.5x showing presumptive synapse. Calibration bar: 5 μm. Cells from undercut and naïve control perfusion-fixed rats are shown in Fig. 3.