Expression and Function of MAO A in Cardiac Cells by Means of Adenovirus-Mediated Gene Transfer
- PMID: 36169862
- DOI: 10.1007/978-1-0716-2643-6_12
Expression and Function of MAO A in Cardiac Cells by Means of Adenovirus-Mediated Gene Transfer
Abstract
Gene-transfer methods are useful to study the structural or functional roles of recombinant proteins in vitro. In particular, adenovirus-mediated gene transduction results in strong efficiency and high level of expression in primary cells such as cardiomyocytes, which are difficult to transfect with classical methods. Here, we describe a protocol that enables efficient expression of MAO A in both primary cells and cell lines. Following expression of recombinant MAO A, substrate-induced activation of the enzyme can be assessed by measuring production of reactive oxygen species and downstream signal transduction pathways in cells. This model allows to decipher the biological function of MAO A on metabolism, mitochondrial fitness, cell death/survival, and proliferation.
Keywords: Adenovirus vector; Cardiomyocyte; Monoamine oxidase; Overexpression; Reactive oxygen species.
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
References
-
- Lang SE, Westfall MV (2015) Gene transfer into cardiac myocytes. Methods Mol Biol 1299:177–190 - DOI
-
- Vannucci L, Lai M, Chiuppesi F, Ceccherini-Nelli L, Pistello M (2013) Viral vectors: a look back and ahead on gene transfer technology. New Microbiol 36:1–22 - PubMed
-
- Lee CS, Bishop ES, Zhang R, Yu X, Farina EM, Yan S, Zhao C, Zheng Z, Shu Y, Wu X, Lei J, Li Y, Zhang W, Yang C, Wu K, Wu Y, Ho S, Athiviraham A, Lee MJ, Wolf JM, Reid RR, He TC (2017) Adenovirus-mediated gene delivery: potential applications for gene and Cell-based therapies in the new era of personalized medicine. Genes Dis 4:43–63 - DOI
-
- Manzella N, Santin Y, Maggiorani D, Martini H, Douin-Echinard V, Passos JF, Lezoualc'h F, Binda C, Parini A, Mialet-Perez J (2018) Monoamine oxidase-A is a novel driver of stress-induced premature senescence through inhibition of parkin-mediated mitophagy. Aging Cell 17:e12811 - DOI
-
- Santin Y, Sicard P, Vigneron F, Guilbeau-Frugier C, Dutaur M, Lairez O, Couderc B, Manni D, Korolchuk VI, Lezoualc'h F, Parini A, Mialet-Perez J (2016) Oxidative stress by monoamine oxidase-A impairs transcription factor EB activation and autophagosome clearance, leading to cardiomyocyte necrosis and heart failure. Antioxid Redox Signal 25:10–27 - DOI
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources