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Moyamoya Disease is two to four times more common in females than males. The underlying mechanisms behind this are currently unknown. Missense mutations in Acta2 predispose affected individuals to a variety of vascular diseases, including Moyamoya-like cerebrovascular disease. In this study we examined vessel wall thickness on H&E stained brain sections from WT and Acta2-/- male and female mice at 12 weeks of age (n=3 per a group). We found that female Acta2-/- mice had significantly greater percent vessel wall thickness compared to male Acta2-/- mice across all vessel sizes. There was not a significant difference between male and female vessel wall thickness in the wild-type groups. These findings in Acta2-/- mice suggest important sex-dependent differences in the function of α-smooth muscle actin (SMA) in the cerebrovascular system, and likely the vascular system as a whole. This has important implications for the design of studies examining the role of SMA in cerebrovascular disease and the investigation of novel therapies.Figure. Percent wall thickness (%TH) by sex. Two-tailed two-sample t-Test assuming unequal variances: * = p<0.05, ** = p<0.005, *** = p<0.0005 and # = one-tailed t-test significant but not two-tailed. SL=short vessel diameter.