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.
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Rassi A, Rassi A, Marin-Neto JA (2010) Chagas disease. Lancet 375:1388–1402
Rassi A, Rassi A, Marcondes de Rezende J (2012) American trypanosomiasis (Chagas Disease). Infect Dis Clin N Am 26:275–291
Cunha-Neto E, Chevillard C (2014) Chagas disease cardiomyopathy: immunopathology and genetics. Mediat Inflamm 2014:683230. https://doi.org/10.1155/2014/683230
Navarro IC, Ferreira FM, Nakaya HI et al (2015) MicroRNA transcriptome profiling in heart of Trypanosoma cruzi-infected mice: parasitological and cardiological outcomes. PLoS Negl Trop Dis 9:e0003828. https://doi.org/10.1371/journal.pntd.0003828
Lee RC, Feinbaum RL, Ambros V (1993) The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 75:843–854. https://doi.org/10.1016/0092-8674(93)90529-Y
Slack FJ, Basson M, Liu Z, Ambros V, Horvitz HR, Ruvkun G (2000) The lin-41 RBCC gene acts in the C. elegans heterochronic pathway between the let-7 regulatory RNA and the LIN-29 transcription factor. Mol Cell 5:659–669. https://doi.org/10.1016/S1097-2765(00)80245-2
Ambros V (2001) microRNAs. Cell 107:823–826. https://doi.org/10.1016/S0092-8674(01)00616-X
Ambros V (2001) microRNAs: tiny regulators with great potential. Cell 107:823–826. https://doi.org/10.1016/S0092-8674(01)00616-X
Ha M, Kim VN (2014) Regulation of microRNA biogenesis. Nat Rev Mol Cell Biol 15:509–524. https://doi.org/10.1038/nrm3838
Krol J, Loedige I, Filipowicz W (2010) The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet 11:597–610. https://doi.org/10.1038/nrg2843
Zhao Y, Ransom JF, Li A, Vedantham V, von Drehle M, Muth AN, Tsuchihashi T, McManus MT, Schwartz RJ, Srivastava D (2007) Dysregulation of cardiogenesis, cardiac conduction, and cell cycle in mice lacking miRNA-1-2. Cell 129:303–317. https://doi.org/10.1016/j.cell.2007.03.030
Carè A, Catalucci D, Felicetti F et al (2007) MicroRNA-133 controls cardiac hypertrophy. Nat Med 13:613–618. https://doi.org/10.1038/nm1582
Boštjančič E, Zidar N, Štajer D, Glavač D (2010) MicroRNAs miR-1, miR-133a, miR-133b and miR-208 are dysregulated in human myocardial infarction. Cardiology 115:163–169. https://doi.org/10.1159/000268088
Duisters RF, Tijsen AJ, Schroen B et al (2009) MiR-133 and miR-30 regulate connective tissue growth factor: Implications for a role of micrornas in myocardial matrix remodeling. Circ Res 104:170–178. https://doi.org/10.1161/CIRCRESAHA.108.182535
Ferreira LRP, Frade AF, Santos RHB et al (2014) MicroRNAs miR-1, miR-133a, miR-133b, miR-208a and miR-208b are dysregulated in chronic Chagas disease cardiomyopathy. Int J Cardiol 175:409–417. https://doi.org/10.1016/j.ijcard.2014.05.019
Tian T, Wang J, Zhou X (2015) A review: microRNA detection methods. Org Biomol Chem 13:2226–2238. https://doi.org/10.1039/c4ob02104e
Kim VN (2005) MicroRNA biogenesis: coordinated cropping and dicing. Nat Rev Mol Cell Biol 6:376–385. https://doi.org/10.1038/nrm1644
Brener Z (1962) Therapeutic activity and criterion of cure on mice experimentally infected with Trypanosoma cruzi. Rev Inst Med Trop Sao Paulo 4:389–396. http://www.imt.usp.br/wp-content/uploads/revista/vol04/389-396.pdf
Gibson UE, Heid CA, Williams PM (1996) A novel method for real time quantitative RT-PCR. Genome Res 6:995–1001. https://doi.org/10.1101/gr.6.10.995
Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Ser B 57:289–300. https://doi.org/10.2307/2346101
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Ferreira, L.R.P. (2019). MicroRNA Transcriptome Profiling in Heart of Trypanosoma cruzi-Infected Mice. In: Gómez, K., Buscaglia, C. (eds) T. cruzi Infection. Methods in Molecular Biology, vol 1955. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9148-8_15
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DOI: https://doi.org/10.1007/978-1-4939-9148-8_15
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