Studying β1 and β2 adrenergic receptor signals in cardiac cells using FRET-based sensors

Publication date: Available online 29 June 2019Source: Progress in Biophysics and Molecular BiologyAuthor(s): Francesca Grisan, Alex Burdyga, Liliana F. Iannucci, Nicoletta C. Surdo, Tullio Pozzan, Giulietta Di Benedetto, Konstantinos LefkimmiatisAbstractCyclic 3′-5′ adenosine monophosphate (cAMP) is a key modulator of cardiac function. Thanks to the sophisticated organization of its pathway in distinct functional units called microdomains, cAMP is involved in the regulation of both inotropy and chronotropy as well as transcription and cardiac death. While visualization of cAMP microdomains can be achieved thanks to cAMP-sensitive FRET-based sensors, the molecular mechanisms through which cAMP-generating stimuli are coupled to distinct functional outcomes are not well understood. One possibility is that each stimulus activates multiple microdomains in order to generate a spatiotemporal code that translates into function. To test this hypothesis here we propose a series of experimental protocols that allow to simultaneously follow cAMP or Protein Kinase A (PKA)-dependent phosphorylation in different subcellular compartments of living cells. We investigate the responses of β Adrenergic receptors (β1AR and β2AR) challenged with selective drugs that enabled us to measure the actions of each receptor independently. At the whole cell level, we used a combination of co-culture with selective βAR stimulation and were able to molecularly separate cardiac fibroblasts from neona...
Source: Progress in Biophysics and Molecular Biology - Category: Molecular Biology Source Type: research