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Published on: September 16, 2011
Characterization of microRNA profile in IgE-mediated mouse BMMCs degranulation
Yanhong Li1, Jie Liu1, Jiaojiao Zhang1
1College of Animal Science and Technology, Southwest University, Chongqing 400715, China.
Background:
Mast cells play a central role in innate and adaptive immunity by releasing pre-formed and de novo synthesized mediators, which include microRNAs. Although miRNAs have been confirmed to function in cell proliferation, differentiation, apoptosis, and the immune response, their functions are still limited in mast cells degranulation.
Methods:
Here, we survey miRNA expression profiles in activated mouse bone marrow-derived mast cells (BMMCs) with a miRNA microarray and compare the profiles to those from resting BMMCs. Partial miRNAs were selected for confirmation by qPCR, and let-7i was selected for function discover in mast cell degranulation process. TargetScan Mouse database were used for target genes prediction, gene ontology (GO) were used for gene molecular function classifications, and Cytoscape software were used to construct gene network of degranulation.
Results:
We found 13 up-regulated miRNAs and 7 down-regulated miRNAs in DNP activated BMMCs by miRNA microarray; and let-7b, let-7c, let-7d, let-7f, let-7i, and miR-652 were up-regulated, and miR-296-3p was down-regulated in DNP-stimulated BMMCs by qPCR. In the function research, let-7i can inhibit mast cell degranulation by suppress Exco8 expression. Overall, the data indicate that miRNAs participate in mast cell activation, especial for mast cell degranulation process.
Insights
MicroRNAs (miRNAs) regulate immune responses. This study reveals that let-7i inhibits mast cell degranulation by targeting Exco8, highlighting miRNAs
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Mast cells are key immune cells releasing mediators, including microRNAs (miRNAs).
- While miRNAs regulate various cellular processes, their role in mast cell degranulation remains underexplored.
Purpose of the Study:
- To investigate miRNA expression profiles in activated mouse bone marrow-derived mast cells (BMMCs).
- To elucidate the function of specific miRNAs, particularly let-7i, in mast cell degranulation.
Main Methods:
- miRNA microarray analysis of activated vs. resting BMMCs.
- Quantitative PCR (qPCR) for miRNA validation.
- Target gene prediction using TargetScan Mouse and gene ontology (GO) analysis.
- Functional studies of let-7i in mast cell degranulation.
Main Results:
- Identified 13 up-regulated and 7 down-regulated miRNAs in activated BMMCs via microarray.
- Confirmed up-regulation of let-7b, let-7c, let-7d, let-7f, let-7i, and miR-652, and down-regulation of miR-296-3p by qPCR.
- Demonstrated that let-7i inhibits mast cell degranulation by suppressing Exco8 expression.
Conclusions:
- MicroRNAs are involved in mast cell activation and degranulation.
- let-7i plays an inhibitory role in mast cell degranulation through Exco8 regulation.
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