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Tissue-specific miRNA Expression Profiling in Mouse Heart Sections Using In Situ Hybridization
Published on: September 15, 2018
Adenosine 2B receptor expression is post-transcriptionally regulated by microRNA
Vasantha L Kolachala1, Lixin Wang, Tracy S Obertone
1Division of Digestive Diseases, Emory University School of Medicine, Atlanta, Georgia 30322, USA. vkolach@emory.edu
The Journal of Biological Chemistry
|April 15, 2010
Summary
Tumor necrosis factor alpha (TNF-alpha) increases adenosine 2B receptor (A(2B)AR) in colitis. This occurs post-transcriptionally, regulated by miR27b and miR128a, with miR27b impacting A(2B)AR in mouse colitis.
Area of Science:
- Molecular Biology
- Immunology
- Gastroenterology
Background:
- Epithelial adenosine 2B receptor (A(2B)AR) mRNA and protein are elevated in colitis.
- Tumor necrosis factor alpha (TNF-alpha) regulates A(2B)AR expression during colitis.
Purpose of the Study:
- To investigate the post-transcriptional mechanisms controlling A(2B)AR expression in colonic epithelial cells during colitis.
- To elucidate the role of microRNAs (miRNAs) in TNF-alpha-mediated A(2B)AR regulation.
Main Methods:
- Cloning of the A(2B)AR promoter and 3'-untranslated region (UTR) into luciferase reporter vectors.
- Utilizing anti-miRNA and miRNA overexpression to assess miRNA-A(2B)AR interactions.
- Quantitative analysis of mRNA levels and promoter activity in T84 cells and mouse colon tissue.
- Nuclear run-on assays to evaluate transcriptional regulation.
Main Results:
- TNF-alpha significantly increased A(2B)AR mRNA but not promoter activity in T84 cells.
- Four potential miRNA target sites (miR27a, miR27b, miR128a, miR128b) were identified in the A(2B)AR 3'-UTR.
- TNF-alpha treatment decreased miR27b and miR128a levels, while their overexpression reduced A(2B)AR expression.
- In vitro blockade of miR27b elevated A(2B)AR mRNA, and miR27b levels were decreased in colitis-affected mouse colon.
Conclusions:
- TNF-alpha-induced A(2B)AR expression in colonic epithelial cells is primarily regulated post-transcriptionally by miR27b and miR128a.
- miR27b plays a significant role in modulating A(2B)AR expression during murine colitis.
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