Fluorescence spectroscopy of low-level endogenous β-adrenergic receptor expression at the plasma membrane of differentiating human iPSC-derived cardiomyocytes
Abstract
The potential of human-induced pluripotent stem cells (hiPSCs) to be differentiated into cardiomyocytes (CMs) mimicking adult CMs functional morphology, marker genes and signaling characteristics has been investigated since over a decade. The evolution of the membrane localization of CM-specific G protein-coupled receptors throughout differentiation has received, however, only limited attention to date. We employ here advanced fluorescent spectroscopy, namely linescan Fluorescence Correlation Spectroscopy (FCS), to observe how the plasma membrane abundance of the β1- and β2-adrenergic receptors (β1/2-ARs), labelled using a bright and photostable fluorescent antagonist, evolves during the long-term monolayer culture of hiPSC-derived CMs. We compare it to the kinetics of observed mRNA levels in wildtype (WT) hiPSCs and in two CRISPR/Cas9 knock-in clones. We conduct these observations against the backdrop of our recent report of cell-to-cell expression variability, as well as of the subcellular localization heterogeneity of β-ARs in adult CMs.
Citation
Gmach , P , Bathe-Peters , M , Telugu , N , Miller , D C & Annibale , P 2022 , ' Fluorescence spectroscopy of low-level endogenous β-adrenergic receptor expression at the plasma membrane of differentiating human iPSC-derived cardiomyocytes ' , International Journal of Molecular Sciences , vol. 23 , no. 18 , 10405 . https://doi.org/10.3390/ijms231810405
Publication
International Journal of Molecular Sciences
Status
Peer reviewed
ISSN
1661-6596Type
Journal article
Description
Funding: This project was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) through Project 421152132 SFB1423 subproject C03 (PA) and SFB 1470 subproject A01 (PA).Collections
Items in the St Andrews Research Repository are protected by copyright, with all rights reserved, unless otherwise indicated.