Live Cell Imaging of Cyclic Nucleotides in Human Cardiomyocytes.
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Live Cell Imaging of Cyclic Nucleotides in Human Cardiomyocytes. / Beneke, Kira; Molina, Cristina.
cAMP Signaling: Methods and Protocols. ed. / Manuela Zaccolo. 2. ed. Methods in Molecular Biology : Humana Press, 2022. p. 195-204 (Methods in Molecular Biology; Vol. 2483).Research output: SCORING: Contribution to book/anthology › Chapter › Research › peer-review
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TY - CHAP
T1 - Live Cell Imaging of Cyclic Nucleotides in Human Cardiomyocytes.
AU - Beneke, Kira
AU - Molina, Cristina
PY - 2022
Y1 - 2022
N2 - The ubiquitous second messengers' 3',5'-cyclic adenosine monophosphate (cAMP ) and 3',5'-cyclic guanosine monophosphate (cGMP) are crucial in regulating cardiomyocyte function, as well as pathological processes, by acting in distinct subcellular microdomains and thus controlling excitation-contraction coupling. Spatio-temporal intracellular dynamics of cyclic nucleotides can be measured in living cells using fluorescence resonance energy transfer (FRET ) by transducing isolated cells with genetically encoded biosensors. While FRET experiments have been regularly performed in cardiomyocytes from different animal models, human-based translational experiments are very challenging due to the difficulty to culture and transduce adult human cardiomyocytes. Here, we describe a technique for obtaining human atrial and ventricular myocytes which allows to keep them alive in culture long enough to transduce them and visualize cAMP and cGMP in physiological and pathological human settings.Keywords: Atrial myocytes; Biosensors; Culture; Fluorescence resonance energy transfer; Human; Isolation; Ventricular myocytes; cAMP; cGMP.
AB - The ubiquitous second messengers' 3',5'-cyclic adenosine monophosphate (cAMP ) and 3',5'-cyclic guanosine monophosphate (cGMP) are crucial in regulating cardiomyocyte function, as well as pathological processes, by acting in distinct subcellular microdomains and thus controlling excitation-contraction coupling. Spatio-temporal intracellular dynamics of cyclic nucleotides can be measured in living cells using fluorescence resonance energy transfer (FRET ) by transducing isolated cells with genetically encoded biosensors. While FRET experiments have been regularly performed in cardiomyocytes from different animal models, human-based translational experiments are very challenging due to the difficulty to culture and transduce adult human cardiomyocytes. Here, we describe a technique for obtaining human atrial and ventricular myocytes which allows to keep them alive in culture long enough to transduce them and visualize cAMP and cGMP in physiological and pathological human settings.Keywords: Atrial myocytes; Biosensors; Culture; Fluorescence resonance energy transfer; Human; Isolation; Ventricular myocytes; cAMP; cGMP.
U2 - 10.1007/978-1-0716-2245-2_12
DO - 10.1007/978-1-0716-2245-2_12
M3 - Chapter
SN - 978-1-0716-2244-5
T3 - Methods in Molecular Biology
SP - 195
EP - 204
BT - cAMP Signaling
A2 - Zaccolo, Manuela
PB - Humana Press
CY - Methods in Molecular Biology
ER -