Figure 3.

RAGE expression in human spermatozoa is linked to multiple markers of poor sperm health. Representative flow cytometry data and quantitative analysis of RAGE positive and negative populations relative to (a and b) MMP as measured by MitoTracker CMXRos, (c–e) apoptosis or necrosis as measured by AnnexinV–DAPI and (f and g) cell permeability as measured by DAPI fluorescence in washed samples, n = 35; % mean ± SEM. Paired t-test; ****P < 0.0001. (h) Correlation between low MMP and RAGE expression in washed sperm samples, n = 35, 95% CI shown by dashed line. ****P < 0.0001. (i) DNA fragmentation defined as a proportion of TUNEL positive spermatozoa in the RAGE positive and negative subgroups, n = 8; % mean ± SEM. Paired t-test; ***P < 0.001. (j) Correlation between TUNEL positive sperm and RAGE expressing sperm, n = 8, 95% CI shown by dashed line. *P < 0.05. (k) Oxidative DNA damage as determined by the number of 8-HOdG positive spermatozoa levels in RAGE positive and RAGE negative subgroups, n = 8, % mean ± SEM. Paired t-test, ***P < 0.001. MMP, mitochondrial membrane potential, 8-HOdG, 8-hydroxy-2-deoxyguanosine.