Dynamic MicroRNA Expression Profiles During Embryonic Development Provide Novel Insights Into Cardiac Sinus Venosus/Inflow Tract Differentiation

Abstract

MicroRNAs have been explored in different organisms and are involved as molecular switches modulating cellular specification and differentiation during the embryonic development, including the cardiovascular system. In this study, we analyze the expression profiles of different microRNAs during early cardiac development. By using whole mount in situ hybridization in developing chick embryos, with microRNA-specific LNA probes, we carried out a detailed study of miR-23b, miR-130a, miR-106a, and miR-100 expression during early stages of embryogenesis (HH3 to HH17). We also correlated those findings with putative microRNA target genes by means of mirWalk and TargetScan analyses. Our results demonstrate a dynamic expression pattern in cardiac precursor cells from the primitive streak to the cardiac looping stages for miR-23b, miR-130a, and miR-106a. Additionally, miR-100 is later detectable during cardiac looping stages (HH15-17). Interestingly, the sinus venosus/inflow tract was shown to be the most representative cardiac area for the convergent expression of the four microRNAs. Through in silico analysis we revealed that distinct Hox family members are predicted to be targeted by the above microRNAs. We also identified expression of several Hox genes in the sinus venosus at stages HH11 and HH15. In addition, by means of gain-of-function experiments both in cardiomyoblasts and sinus venosus explants, we demonstrated the modulation of the different Hox clusters, Hoxa, Hoxb, Hoxc, and Hoxd genes, by these microRNAs. Furthermore, we correlated the negative modulation of several Hox genes, such as Hoxa3, Hoxa4, Hoxa5, Hoxc6, or Hoxd4. Finally, we demonstrated through a dual luciferase assay that Hoxa1 is targeted by miR-130a and Hoxa4 is targeted by both miR-23b and miR-106a, supporting a possible role of these microRNAs in Hox gene modulation during differentiation and compartmentalization of the posterior structures of the developing venous pole of the heart.

miR-195b is required for proper cellular homeostasis in the elderly

“…Embryos were collected at distinct embryonic stages as previously described, randing from E10-5 to E16.5, fixed for 2 h at 4 °C in 4% PFA, dehydrated in graded ethanol steps and stored at − 20 °C. Embryos were processed for whole-mount LNA-ISH was processed as previously described 33 . miR-195 LNA-labeled microRNA probe (miRCURY LNA™ Detection probe 5′-DIG and 3′-DIG labeled, Exiqon) was used to performed whole-mount LNA ISH.…”

Identification of circRNA–miRNA–mRNA Regulatory Network and Crucial Signaling Pathway Axis Involved in Tetralogy of Fallot

“…Similarly, hsa-miR-181b has been found to correlate with hypoxia-triggered cardiomyocyte apoptosis ( Lv et al, 2020 ). Hsa-miR-130a affects cardiac sinus venous and inflow tract differentiation ( Garcia-Padilla et al, 2021 ). In summary, we speculate that these miRNAs are mainly involved in the division, differentiation, and apoptosis of cardiomyocytes during the pathogenesis of TOF.…”

Comparative Analysis of Non-Coding RNA Transcriptomics in Heart Failure

“…MicroRNAs are on average 20–22 ribonucleotides in length and display the capacity to bind to the 3’ untranslated region (3´UTR) of coding RNAs by complementary base pairing, promoting their degradation and/or translational blockage. The role of microRNAs as post-transcriptional modulators has been widely described in multiple biological processes, including cell development, differentiation, growth and homeostasis [ 60 , 61 , 62 , 63 ]. Interestingly, it has been widely described that microRNAs are associated with cardiac physiological and pathological phenotypes, and nowadays circulating microRNAs are being deeply studied as potential HF diagnostic biomarkers.…”