Figure 8
Differences between capillary rarefaction, hypertrophy and overall cardiac disorganization in ZDF-F compared to ZDF-M rats. (a) Graph shows heart weight adjusted to tibia length indicating ZDF-F heart is hypertrophied compared to ZL-F. (b) Quantification of cardiomyocyte cross-sectional area determined from HPA-staining (c) Quantification of capillary density determined from IB4-staining. (d) Representative images of heart sections stained with HPA conjugated to Alexa Fluor 647 to visualize cardiomyocyte membrane and IB4 conjugated to Alexa Fluor 594 to visualize capillaries (scale bars = 75 µm); n = 5–8 animals per group. For cardiomyocyte hypertrophy calculations, number of cardiomyocytes analyzed ranged from 171 to 181 per animal. Values are means ± SEM. p-values are noted and were determined by two-way ANOVA. *p < 0.05 vs. ZL-F, †p < 0.05 vs. ZL-M. ZDF-F cardiomyocytes were significantly larger than any of the other 3 groups (p < 0.05). (e) Representative immunoblot that shows the extent of Ser2448phosphorylation levels of mTOR protein. Graph shows quantification of Ser2448phosphorylation of mTOR. n = 6 animals per group. (f) Representative images of cardiac tissues stained with Masson’s trichrome (MTS). Only ZDF-F exhibited scar tissue (marked with yellow arrow). (g) Representative images of cardiac tissues stained and hematoxylin and eosin (H&E). Yellow arrows mark blood cells spilling into interstitial regions in ZDF-F. Scale bars = 100 µm. n = 5–8 animals per group. (h) Representative transmission electron micrographs at 1000X showing extensive disruption of the normal cardiac organization in ZDF-M and ZDF-F compared with respective leans. n = 3–5 animals per group. Yellow scale bars = 2 µm.
