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Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs
Published on: June 23, 2022
MicroRNA Transcriptome Profiling in Heart of Trypanosoma cruzi-Infected Mice
Ludmila Rodrigues Pinto Ferreira1
1Departamento de Morfologia, RNA Systems Biology Laboratory (RSBL), Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Minas Gerais, Brazil. ludmila@icb.ufmg.br.
Abstract:
MicroRNAs (miRNAs) are a class of small noncoding RNAs (typically 19-23 nucleotides) which act by annealing to partially complementary binding sites present on the 3' untranslated regions (UTR) of messenger RNAs (mRNAs) leading to inhibition of protein translation or by inducing mRNA decay. Since their discovery, miRNAs have come to be recognized as master regulators of gene expression in plant and mammals, controlling tissue-specific protein expression. Up to one-third of mammalian mRNAs are susceptible to miRNA-mediated regulation. It has been shown that miRNAs are determinants of the physiology and pathophysiology of the cardiovascular system, and altered expression of muscle- and/or cardiac-specific miRNAs in myocardial tissue is involved in heart development and cardiovascular diseases, including myocardial hypertrophy, heart failure, and fibrosis. The analysis of miRNA expression pattern provides important information, as well as is a starting point to understand miRNA function in different tissues, during development, and in disease. Several techniques can be used for miRNA profiling analysis like high-throughput sequencing, microarrays, and real-time PCR using microfluidic low-density arrays. This chapter describes the complete methodology to perform miRNA profiling using the stem-loop reverse-transcription (RT)-based TaqMan® MicroRNA low-density arrays (TLDA) method. This methodology was used to perform miRNA profiling in the heart of T. cruzi acutely infected mice.
Insights
MicroRNAs regulate gene expression and are vital in cardiovascular health and disease. This study details a method for profiling microRNA expression in heart tissue, specifically in mice infected with T. cruzi.
Area of Science:
- Molecular Biology
- Genetics
- Cardiovascular Research
Background:
- MicroRNAs (miRNAs) are small noncoding RNAs regulating gene expression in mammals, affecting up to one-third of mRNAs.
- Dysregulated miRNA expression in cardiac tissue is implicated in cardiovascular diseases like hypertrophy, heart failure, and fibrosis.
- Understanding miRNA function requires analyzing their expression patterns in various physiological and pathological conditions.
Purpose of the Study:
- To describe a comprehensive methodology for microRNA profiling.
- To apply this method for analyzing miRNA expression in the heart tissue of mice during acute T. cruzi infection.
Main Methods:
- Utilized the stem-loop reverse-transcription (RT)-based TaqMan® MicroRNA low-density arrays (TLDA) for miRNA profiling.
- Detailed the complete methodology for performing miRNA profiling using TLDA.
- Applied the methodology to profile miRNAs in the heart of T. cruzi acutely infected mice.
Main Results:
- Successfully profiled microRNA expression in the heart tissue of T. cruzi acutely infected mice using the TLDA method.
- Established a complete workflow for miRNA profiling applicable to cardiovascular research.
- Provided a foundation for understanding miRNA roles in T. cruzi-induced cardiac pathology.
Conclusions:
- The described TLDA-based miRNA profiling methodology is effective for analyzing cardiac miRNA expression.
- This approach is valuable for investigating the role of miRNAs in cardiovascular diseases, including infectious etiologies like T. cruzi infection.
- The study provides insights into miRNA expression changes in the context of acute Chagas disease.
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