Figure 1.
Cross-species microRNA/mRNA cardiac atlas.
Data sets obtained for rat, dog and cynomolgus monkey are indicated. Inset: magnified view of a papillary muscle.
Figure 2.
Distribution of microRNAs and mRNAs in rat cardiac structures.
(A) Cardiac samples are grouped according to the structure using microRNA signatures (top 10%) in a hierarchical Euclidean-linkage clustering based on miRBase17 mapping (B) Principal Component Analysis of all microRNA profiles based on miRBase17 mapping. (C) Principal Component Analysis of all mRNA profiles. Red circles: myocardial tissue (apex, left and right ventricle, septum, papillary muscle). Blue circles: left and right atrium. Grey circles: cardiac valves. A, apex. LA, left atrium. LV, left ventricle. PM, papillary muscle. RA, right atrium. RV, right ventricle. S, septum. V, valves. Individual animals are indicated by symbols.
Figure 3.
Number of microRNAs (A) and mRNAs (B) preferentially expressed in cardiac valves or myocardium across species.
Myocardium includes apex, ventricles, septum and papillary muscles. Fold change thresholds were > = 2 for microRNAs in (A), and > = 5 from mRNAs in (B).
Figure 4.
Distribution of miR-1, miR-125b-5p, miR-204 and miR-208b in cardiac structures across species.
Axes represent fold change vs. apex. A, apex; LA, left atrium; LV, left ventricle; PM, papillary muscle; RA, right atrium; RV, right ventricle; S, septum; V, valve.
Figure 5.
Localization of miR-204, miR-125b-5p, miR-1 and miR-122 in rat heart by in situ hybridization.
miR-204 in valves (A–B) and ventricle (C). miR-125b-5p in valves (D–E) and ventricle (F). miR-1 in valves (G–H) and ventricle (I). miR-122 in valves (J–K) and ventricle (L). Bar = 100 µm.
Figure 6.
Negative correlation of cardiac tissue microRNA and target mRNA profiles.
Timp3 and miR-1 (A), Rbm24 and miR-125b-5p (B), Tgfbr2 and miR-204 (C), Csnk2a2 and miR-208b (D). Green curve represents log2 normalized intensity for the indicated probe set. Black curve represents log2 scaled and normalized microRNA read counts. Three replicates are plotted for each structure. A, apex; LA, left atrium; LV, left ventricle; PM, papillary muscle; RA, right atrium; RV, right ventricle; S, septum; VM, valve.
Figure 7.
Anti-correlated microRNA targets are directly inhibited by microRNA over expression.
(A–D) Real-Time RT-PCR of Timp3, Rbm24, Tgfbr2 and Csnk2a2 in HPASM cells transfected with mimics for miR-1, miR-125b-5p, miR-204, miR-499 and miR-208b or with a mimic microRNA negative control. Data were normalized to 18S RNA. (E-H) Luciferase activity of wild-type (WT) or mutant (MUT) Timp3, Rbm24, Tgfbr2 and Csnk2a2 3′-UTR reporter genes cotransfected with their paired microRNA mimics. Data were averaged for n = 4 in 3 independent experiments and error bars represented standard deviation of the mean. P values: * = P<0.05, ** = P<0.01, *** = P<0.005.