Fluorescence spectroscopy of low-level endogenous β-adrenergic receptor expression at the plasma membrane of differentiating human iPSC-derived cardiomyocytes
Please always quote using this URN: urn:nbn:de:bvb:20-opus-288277
- 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 plasmaThe 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.…
Author: | Philipp Gmach, Marc Bathe-Peters, Narasimha Telugu, Duncan C. Miller, Paolo Annibale |
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URN: | urn:nbn:de:bvb:20-opus-288277 |
Document Type: | Journal article |
Faculties: | Medizinische Fakultät / Institut für Pharmakologie und Toxikologie |
Language: | English |
Parent Title (English): | International Journal of Molecular Sciences |
ISSN: | 1422-0067 |
Year of Completion: | 2022 |
Volume: | 23 |
Issue: | 18 |
Article Number: | 10405 |
Source: | International Journal of Molecular Sciences (2022) 23:18, 10405. https://doi.org/10.3390/ijms231810405 |
DOI: | https://doi.org/10.3390/ijms231810405 |
Dewey Decimal Classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
Tag: | CRISPR/Cas9; FCS; GPCR; cardiomyocyte; differentiation; fluorescence; fluorescence correlation spectroscopy; hiPSC-CM; β-adrenergic receptors |
Release Date: | 2023/09/05 |
Date of first Publication: | 2022/09/08 |
Licence (German): | CC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International |