Figure 7.

Plasma pro-inflammatory markers and spleen size after bilateral CSDs. (a) CSDs were induced either by optogenetics or by brief topical KCl application through thinned skull, and monitored by laser Doppler flowmetry. Six CSDs were induced in each hemisphere. In one subset (left panel), all six CSDs were induced in one hemisphere over an hour and then this was repeated in the other hemisphere. In the second subset (right panel), CSDs were induced bilaterally at the same time over an hour. Plasma was collected at 1, 4, or 6 h. Spleen was harvested at 4 or 18–24 h after the first CSD and weighed. Data from bilateral CSD animals were compared with sham controls. (b) Plasma inflammatory markers (IL-1β, CCL2, TNF-α, IL-6, ICAM-1, and VCAM-1) at 1, 4, or 6 h after first CSD or sham (whiskers, full range; box, interquartile range; horizontal line, median; + , mean). Individual data points from each animal are also shown. Dotted lines represent lower limit of reliable quantification based on the standard curves. There was no difference between sham and CSD group in any of the markers at any time point (unpaired t-test). (c) Spleen size in naïve animal, 4 or 18–24 h after first CSD or sham, or 24 h after middle cerebral artery occlusion (stroke). There was no significant difference between sham and CSD group within each time point. Stroke animal showed significant decrease compared to naïve animal. * – *** p < 0.05–0.001 vs. naïve (one-way ANOVA followed by Tukey's multiple comparisons test). LDF: laser Doppler flowmetry; CSD: cortical spreading depolarization; Cox-2: Cyclooxygenase-2; IL-1β: interleukin-1β; CCL2: chemokine (C-C motif) ligand 2; TNF-α: tumor necrosis factor-α; IL-6: interleukin-6; ICAM-1: intercellular adhesion molecule-1; VCAM-1: vascular cell adhesion molecule-1.