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Fluorescence spectroscopy of low-level endogenous ß-adrenergic receptor expression at the plasma membrane of differentiating human iPSC-derived cardiomyocytes

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Item Type:Article
Title:Fluorescence spectroscopy of low-level endogenous ß-adrenergic receptor expression at the plasma membrane of differentiating human iPSC-derived cardiomyocytes
Creators Name:Gmach, P. and Bathe-Peters, M. and Telugu, N. and Miller, D.C. and Annibale, P.
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 β(1)- and β(2)-adrenergic receptors (β(1/2)-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.
Keywords:GPCR, ß-Adrenergic Receptors, hiPSC-CM, Cardiomyocyte, Fluorescence Correlation, Spectroscopy, FCS, Fluorescence, CRISPR/Cas9, Differentiation
Source:International Journal of Molecular Sciences
ISSN:1422-0067
Publisher:MDPI
Volume:23
Number:18
Page Range:10405
Date:8 September 2022
Official Publication:https://doi.org/10.3390/ijms231810405
PubMed:View item in PubMed

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