Presence of IL6 and IL8 receptors within mesenteric and skeletal muscle microvessels in mice

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Metabolic diseases like inflammatory conditions result in an upregulation of proinflammatory cytokines (Diabetes Research and Clinical Practice: 105(2); 141–50, 2014). We have previously examined vasoactive response to IL6 and IL8 in mesenteric and skeletal muscle microvessels in normal wild‐type C57 BL/6J and leptin‐deficient db/db mice. Due to the apparent cross‐talk with endothelin related vasoactive responses (The FASEB Journal 27: 687.15, 2013), we confirmed the presence of these receptors within the tissues. Receptors were identified using immunofluorescence with specific antibodies directed towards the IL6 (ABCAM ab83053) and IL8 (CXCR2, ab14935) receptors. Mesenteric tissues and skeletal muscle tissues from C57BL/6J mice [obtained through a tissue sharing facility at SBU] were fixed in 4% paraformaldehyde overnight, then in 30% sucrose in saline overnight to 5 days, imbedded in freezing medium (optimal cutting temperature compound, OCT) and kept at −80C until use. Tissues were sliced 10 um thick using a cryotome. Tissues were first incubated at room temperature with blocking buffer (0.2% BSA). Primary antibodies were diluted to 1:50 to 1:200, and tissues incubated for 30–60 minutes, rinsed 3–6 times with PBS, and then incubated with FITC IgG secondary antibodies (1:50–1:200) for 30–60 minutes, with a final wash in PBS (3–6 times). Tissues were mounted with fluormount and kept cool in the dark until imaging. The results showed that in both mesenteric microvessels and skeletal muscle microvessels, there was a punctate appearance along the axial distribution of vessels indicative of positive staining to both IL6 and IL8 receptor antibodies. These results indicate that vasoactive responses to the IL6 and IL8 cytokines may be specific in nature, and not solely through the endothelin receptor system. We have not ruled out a potential interaction between separate receptor systems as being a requirement for vasoactive responses.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.