Energy Metabolism and Intracellular pH Alteration in Neural Spheroids Carrying Down Syndrome

Autor: Alena Kashirina, Alena Gavrina, Emil Kryukov, Vadim Elagin, Yuliya Kolesova, Alexander Artyuhov, Ekaterina Momotyuk, Vepa Abdyyev, Natalia Meshcheryakova, Elena Zagaynova, Erdem Dashinimaev, Aleksandra Kashina
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: Biomedicines, Vol 9, Iss 11, p 1741 (2021)
Druh dokumentu: article
ISSN: 2227-9059
DOI: 10.3390/biomedicines9111741
Popis: Brain diseases including Down syndrome (DS/TS21) are known to be characterized by changes in cellular metabolism. To adequately assess such metabolic changes during pathological processes and to test drugs, methods are needed that allow monitoring of these changes in real time with minimally invasive effects. Thus, the aim of our work was to study the metabolic status and intracellular pH of spheroids carrying DS using fluorescence microscopy and FLIM. For metabolic analysis we measured the fluorescence intensities, fluorescence lifetimes and the contributions of the free and bound forms of NAD(P)H. For intracellular pH assay we measured the fluorescence intensities of SypHer-2 and BCECF. Data were processed with SPCImage and Fiji-ImageJ. We demonstrated the predominance of glycolysis in TS21 spheroids compared with normal karyotype (NK) spheroids. Assessment of the intracellular pH indicated a more alkaline intracellular pH in the TS21 spheroids compared to NK spheroids. Using fluorescence imaging, we performed a comprehensive comparative analysis of the metabolism and intracellular pH of TS21 spheroids and showed that fluorescence microscopy and FLIM make it possible to study living cells in 3D models in real time with minimally invasive effects.
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