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- Title
MiR-132 Regulates Rem Expression in Cardiomyocytes During Long-Term β-Adrenoceptor Agonism.
- Authors
Carrillo, Elba D.; Sampieri, Raúl; Hernández, Ascención; García, María C.; Sánchez, Jorge A.
- Abstract
Aims:To characterize the effects of long-term β-adrenergic receptor stimulation on Rem protein and mRNA expression in rat heart and possible involvement of miR-132. Methods:Adult rats were treated with isoproterenol (ISO, 150 µg.kg.h-1) for 2 d and Rem, miR-132, and α1c (the principal subunit of Cav1.2 channels) were measured at protein and mRNA levels with western blot and quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) experiments, respectively. Ca2+ currents and intracellular Ca2+ signals were evaluated in isolated cardiomyocytes. Results:Systemic administration of ISO led to decreases in Rem protein and mRNA levels (down to 49%). Furthermore, levels of the microRNAs (miRs) miR-132 and miR-214 were upregulated 5- and 9-fold, respectively. Transfection of miR-132, but not miR-214, into HEK293 cells reduced the expression of a luciferase reporter gene controlled by a conserved 3´-untranslated region (UTR) of Rem by half. Chronic ISO administration also led to a 25% decrease in the amplitude of peak L-type Ca2+ currents, a 40% decrease in α1c subunit protein abundance at the membrane level, and a 60% decrease in expression of α1c channel subunit mRNA. Conclusions:These results suggest that Rem expression is down-regulated posttranscriptionally by miR-132 in response to long-term activation of β-adrenergic signaling, but this down-regulation does not produce a larger Ca2+ influx through Cav1.2 channels. © 2015 S. Karger AG, Basel
- Subjects
MICRORNA; GENE expression; GENE transfection; HEART cells; REVERSE transcriptase polymerase chain reaction; ISOPROTERENOL
- Publication
Cellular Physiology & Biochemistry (Karger AG), 2015, Vol 36, Issue 1, p141
- ISSN
1015-8987
- Publication type
Article
- DOI
10.1159/000374059