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. 2016 Nov 30;158(3):440–456. doi: 10.1097/j.pain.0000000000000778

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Copyright © 2016 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the International Association for the Study of Pain

This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (CCBY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Immunostaining for substance P(SP), calcitonin gene-related peptide (CGRP), and vesicular neurotransmitter transporters in lamina II of the dorsal horn. (A) A field that contains several SP-immunoreactive profiles (red). (B) The same field scanned to reveal vesicular glutamate transporter 2 (VG2, blue) and vesicular GABA transporter (VGAT, green). (C) A merged image showing SP, VG2, and VGAT. (D) The same field scanned to reveal SP (red) and CGRP (blue). Comparison of these images shows that some SP-immunoreactive profiles are strongly labelled for VG2 and are likely to correspond to axons of excitatory preprotachykinin A–expressing neurons (one indicated with arrow), whereas others are immunoreactive for CGRP and represent central terminals of peptidergic primary afferents (one shown with double arrowhead). Occasionally, VGAT-positive profiles with very weak SP immunoreactivity are present, and one of these is indicated with an arrowhead. All images are projections of 3 optical sections at 0.3 μm z-spacing. Scale bar = 5 μm.