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. 2011 Apr 13;106(1):44–58. doi: 10.1152/jn.00897.2010

Fig. 5.

Fig. 5.

PIIH-like and synaptotagmin immunoreactivity in the STG. A: anti-PIIH (Ai) and anti-synaptotagmin (Aii) both labeled in the fine neuropil. A colocalization channel was generated and overlaid with the PIIH-like and synaptotagmin-labeled channels to reveal overlap in a single optical section of a sectioned STG (filled arrowheads in Aiii). Notice that the majority of PIIH-like immunolabeled patches did not colocalize with synaptotagmin immunoreactivity (open arrowheads in Aiii); also, many synaptotagmin-positive spots did not overlap with PIIH-like immunolabeling (stars in Aiii). Scale bar, 50 μm. B: PIIH-like and synaptotagmin immunoreactivity on a NB-labeled IC neuron, visualized as 3-D rendered transparency projection from 17 1.3-μm optical slices. Colocalization channels were generated to show only immunolabeling in or on the surface of the filled cell. The yellow colocalization channel (Bi) was generated from intensity correlations between the IC neuropil and PIIH-like immunolabeling, and the blue colocalization channel (Bii) was generated from intensity correlations between the IC neuropil and synaptotagmin immunoreactivity. Overlay of the IC neuropil fill and both colocalization channels reveals PIIH-like immunolabeled branches in yellow (open arrowheads in Biii), patches of colocalization with synaptotagmin immunolabeling in blue (stars in Biii), and triple labeling of PIIH-like and synaptotagmin immunolabel colocalization in white (filled arrowheads in Biii). Scale bar, 40 μm. C: enhanced zoom of boxed area in Biii shows a PIIH-like labeled branch of the IC neuropil with a distinct patch of synaptotagmin colocalization (filled arrowhead in Ci). Overlay of the unprocessed channels (Cii), which also reveals protein outside of the IC neuropil, shows that the PIIH-like signal at this location appears to follow along the IC branch (open arrowhead in Cii) and therefore is likely to be postsynaptic, whereas the synaptotagmin signal shows along a different process (open arrows in Ciii), which forms a close contact with the IC branch, without any indication of presynaptic PIIH-like labeling in this presynaptic process. Scale bar, 13 μm.