Figure 1

Necropsy, histology and lipids of TBI and sham mice. (A) Necropsy of exemplary TBI and sham mice of cohort-1, 101 days after surgery. (B) Immunofluorescence analysis in exemplary TBI and sham mice of cohort-1. The images show the microglial marker, ionized calcium-binding adapter molecule 1 (IBA1) and the astroglial marker, glial fibrillary acidic protein (GFAP) in the perilesional and corresponding contra-lesional/contra-lateral area. The images were captured on an LSM510 confocal microscope (Zeiss) and exported from LSM Image Browser 4.2 to Adobe Photoshop CC2018 for a global adjustment of color balance. Scale bars are 50 µm and 20 µm for upper and bottom panels. The number of IBA1 + and GFAP + cells was counted with the particle counter implemented in FIJI ImageJ (https://imagej.net/Fiji) after threshold settings according the 'auto' detection. The bars show mean and SD, the scatter show individual mice. Asterisks show significant differences versus sham ipsilateral, one-way ANOVA, posthoc Holm Šidák, adjusted ****P = 0.0001, **P 0.0091. (C) Box/scatter plots showing brain endocannabinoid concentrations in ipsilateral perilesional cortex (Ctx) and subcortex (subCtx) and corresponding regions of the contra lateral side 155 days after TBI or sham surgery (cohort-2). The endocannabinoids anandamide (AEA), oleoylethanolamide (OEA), palmitoylethanolamide (PEA) and 1 plus 2 arachidonoylglycerol (1,2-AG) were analyzed per tandem LC–MS/MS. The box shows the interquartile range, the line is the median, the whiskers show minimum to maximum, the scatter are individual mice. Data were compared per 2-way ANOVA ("site" × "group") and subsequent posthoc t test for the between subject factor group, i.e. TBI versus sham. Asterisks show multiplicity adjusted P values (*< 0.05). Graphs were created with Graphpad Prism 8.4 (https://www.graphpad.com), exported as enhanced metafile (emf), and arranged and labeled in Adobe Illustrator CC2020 (https://www.adobe.com/de), and exported to TIFF format.