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Rhodamine-based sensor for real-time imaging of mitochondrial ATP in living fibroblasts

Authorsde la Fuente-Herreruela, D.; Gónzalez-Charro, V.; Almendro-Vedia, V.G.; Morán, M.; Martín, M.Á.; Lillo, M.Pilar; Natale, P.; López-Montero, I.
Issue Date2017
CitationBiochimica et Biophysica Acta - Bioenergetics 1858: 999- 1006 (2017)
AbstractMitochondria are essential for the production and maintenance of ATP in the eukaryotic cell. To image and monitor intracellular ATP level without cell breakage, biological and chemical sensors were developed in the last years. Here, we have internalized a rhodamine-based sensor RSL into living cells and monitored the mitochondrial ATP levels in cultured mouse embryonic fibroblasts. To evaluate the robustness of the sensor we imaged the changes of the mitochondrial ATP levels under non-physiological conditions upon incubation with FCCP, oligomycin, azide, deoxyglucose or phosphoenolpyruvate; all compounds that interfere with ATP homeostasis of the cell. The ATP sensor allowed us to determine the mitochondrial ATP levels in human skin fibroblasts where we observe a similar amount of ATP compared to mouse embryonic fibroblasts. We propose the RSL to be a valuable tool for the assessment of mitochondrial dysfunction in human cells derived from mitochondrial OXPHOS patients and for basic studies on bioenergetics metabolism.
Identifiersdoi: 10.1016/j.bbabio.2017.09.004
issn: 1879-2650
Appears in Collections:(IQFR) Artículos
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