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oar-miR-411a-5p Promotes Proliferation and Differentiation in Hu Sheep Myoblasts Under Heat Stress by Targeting SMAD2
Jiawei Lu1, Yilan Liu1, Huixia Li1
1College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China.
Abstract:
MicroRNAs (miRNAs) are endogenous noncoding RNAs that produce a remarked effect on regulating posttranscriptional gene expression. Our previous study identified a decrease in the expression of oar-miR-411a-5p from umbilical plasma in intrauterine growth restriction (IUGR) Hu lambs subjected to maternal heat stress. In this study, we demonstrated that oar-miR-411a-5p could modulate skeletal muscle development. Overexpression of oar-miR-411a-5p significantly enhanced proliferation and differentiation in heat-stressed Hu sheep myoblasts while suppressing apoptosis. Conversely, inhibition of oar-miR-411a-5p resulted in opposing effects. Subsequently, RNAhybrid analysis revealed targeted sites between oar-miR-411a-5p and the 3' untranslated region (UTR) of SMAD2. This suggested that SMAD2 is a direct target gene of oar-miR-411a-5p, as its expression was negatively modulated by oar-miR-411a-5p, a finding corroborated by dual-luciferase assay and RT-qPCR. Furthermore, co-transfection of oar-miR-411a-5p and SMAD2 into Hu sheep myoblasts indicated that oar-miR-411a-5p modulated heat-stressed myoblast growth by targeting SMAD2. In conclusion, these findings elucidate the function of oar-miR-411a-5p in promoting the development of heat-stressed Hu sheep myoblasts, thereby enhancing our understanding of how miRNAs influence skeletal muscle growth in heat-stressed Hu sheep.
Insights
Oar-miR-411a-5p promotes skeletal muscle development in heat-stressed sheep by targeting SMAD2. This microRNA enhances myoblast proliferation and differentiation, offering insights into heat stress impacts on livestock growth.
Area of Science:
- Molecular Biology
- Animal Science
- Genetics
Background:
- MicroRNAs (miRNAs) regulate gene expression post-transcriptionally.
- Previous work showed decreased oar-miR-411a-5p in intrauterine growth restriction (IUGR) lambs under heat stress.
- Heat stress negatively impacts livestock development and productivity.
Purpose of the Study:
- To investigate the role of oar-miR-411a-5p in skeletal muscle development under heat stress.
- To identify the molecular targets of oar-miR-411a-5p in heat-stressed sheep myoblasts.
Main Methods:
- Overexpression and inhibition of oar-miR-411a-5p in Hu sheep myoblasts.
- RNAhybrid analysis to predict miRNA targets.
- Dual-luciferase assay and RT-qPCR to validate gene targets.
- Co-transfection experiments to confirm functional interactions.
Main Results:
- Oar-miR-411a-5p overexpression enhanced proliferation and differentiation while suppressing apoptosis in heat-stressed myoblasts.
- SMAD2 was identified as a direct target gene of oar-miR-411a-5p.
- Oar-miR-411a-5p regulates heat-stressed myoblast growth by targeting SMAD2.
Conclusions:
- Oar-miR-411a-5p plays a crucial role in promoting skeletal muscle development in heat-stressed Hu sheep.
- This study elucidates a novel miRNA-mediated pathway influencing myoblast growth under thermal stress.
- Findings contribute to understanding miRNA functions in livestock adaptation to environmental challenges.
