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. 2015 May 9;11:26. doi: 10.1186/s12990-015-0024-3

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© Rice et al.; licensee BioMed Central. 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Nav1.7 immunolabeling (IL) of arterioles (Ar), arteriole-venule shunts (AVS) and associated innervation in normal human plantar glabrous skin with Alomone (A, B) or Yale (C) Nav1.7 antibodies (red). Co-labeling of innervation (arrows) as marked with anti-PGP 9.5 (PGP, green, A) or smooth muscle cells in tunica media (tm) as marked with anti α-smooth muscle actin antibody (αSMA, green, B,C). Nuclei are DAPI-labeled (blue). Left images (each panel) show only red fluorescence, middle images green; right images show triple-label combinations. Large white rectangles are 2X-enlargements of small rectangles. A-C. Nav1.7-IL is expressed on endothelial cells of tunica intima (red arrowheads) and tm smooth muscle cells as confirmed by double-labeling with anti-αSMA (B, C). Nav1.7-IL is expressed on virtually all vascular innervation (arrows) in tunica adventitia (ta) as confirmed by anti-PGP 9.5 double-labeling (A, yellow arrows). N=nerve. D-E. Nav1.7-IL on arteriole endothelial cells shown as 2X-enlargements of areas indicated by white rectangles in B,C. First images (each panel) show Nav1.7-IL on smooth muscle cells in tm and endothelial cells (red arrowheads). The second images show α-SMA co-labeling of only the smooth muscle cells of tm (green). The third images show merge of first and second images with DAPI (blue). Sections re-labeled with anti-PECAM (green) to show co-labeling with Nav1.7 on endothelial cells (yellow arrowheads, fourth and fifth images). F-G. Background Cy3 fluorescence is limited with no primary antibody in arteriole deep in dermis (F), epidermis (Ep) and upper-dermis (UD) (G). In F, broken line shows tm perimeter with dotted line around arteriole lumen. In G, broken line indicates basement membrane of epidermis and dotted line indicates boundary of dead and live superficial keratinocyte layers (stratum corneum, sc and stratum granulosum, sg, respectively). Stratum spinosum, ss; stratum basalis, sb; dermal papilla (dp). Scale bars=150μm (A); 100μm (B ,C, F, G); 50μm in D,E.