TY - JOUR
T1 - Quantification of the Metabolic State in Cell-Model of Parkinsona (tm) s Disease by Fluorescence Lifetime Imaging Microscopy
AU - Chakraborty, Sandeep
AU - Nian, Fang Shin
AU - Tsai, Jin Wu
AU - Karmenyan, Artashes
AU - Chiou, Arthur
N1 - Funding Information:
This work has been supported by the Ministry of Science and Technology, Taiwan, ROC (Project No. NSC102-2218-E-010-003) and the Top University Project from Taiwan Ministry of Education, ROC. The authors would also like to acknowledge the support of Prof. Hsin-Chen Lee, Department & Institute of Pharmacology, School of Medicine, National Yang-Ming University, Taipei-112, Taiwan, for generously providing the kits and other resources to perform the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) assays.
PY - 2016/1/13
Y1 - 2016/1/13
N2 - Intracellular endogenous fluorescent co-enzymes, reduced nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD), play a pivotal role in cellular metabolism; quantitative assessment of their presence in living cells can be exploited to monitor cellular energetics in Parkinson's disease (PD), a neurodegenerative disorder. Here, we applied two-photon fluorescence lifetime imaging microscopy (2P-FLIM) to noninvasively measure the fluorescence lifetime components of NADH and FAD, and their relative contributions in MPP + (1-methyl-4-phenylpyridinium) treated neuronal cells, derived from PC12 cells treated with nerve growth factor (NGF), to mimic PD conditions. A systematic FLIM data analysis showed a statistically significant (p < 0.001) decrease in the fluorescence lifetime of both free and protein-bound NADH, as well as free and protein-bound FAD in MPP + treated cells. On the relative contributions of the free and protein-bound NADH and FAD to the life time, however, both the free NADH contribution and the corresponding protein-bound FAD contribution increase significantly (p < 0.001) in MPP + treated cells, compared to control cells. These results, which indicate a shift in energy production in the MPP + treated cells from oxidative phosphorylation towards anaerobic glycolysis, can potentially be used as cellular metabolic metrics to assess the condition of PD at the cellular level.
AB - Intracellular endogenous fluorescent co-enzymes, reduced nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD), play a pivotal role in cellular metabolism; quantitative assessment of their presence in living cells can be exploited to monitor cellular energetics in Parkinson's disease (PD), a neurodegenerative disorder. Here, we applied two-photon fluorescence lifetime imaging microscopy (2P-FLIM) to noninvasively measure the fluorescence lifetime components of NADH and FAD, and their relative contributions in MPP + (1-methyl-4-phenylpyridinium) treated neuronal cells, derived from PC12 cells treated with nerve growth factor (NGF), to mimic PD conditions. A systematic FLIM data analysis showed a statistically significant (p < 0.001) decrease in the fluorescence lifetime of both free and protein-bound NADH, as well as free and protein-bound FAD in MPP + treated cells. On the relative contributions of the free and protein-bound NADH and FAD to the life time, however, both the free NADH contribution and the corresponding protein-bound FAD contribution increase significantly (p < 0.001) in MPP + treated cells, compared to control cells. These results, which indicate a shift in energy production in the MPP + treated cells from oxidative phosphorylation towards anaerobic glycolysis, can potentially be used as cellular metabolic metrics to assess the condition of PD at the cellular level.
UR - http://www.scopus.com/inward/record.url?scp=84955098754&partnerID=8YFLogxK
U2 - 10.1038/srep19145
DO - 10.1038/srep19145
M3 - Article
C2 - 26758390
AN - SCOPUS:84955098754
SN - 2045-2322
VL - 6
JO - Scientific Reports
JF - Scientific Reports
M1 - 19145
ER -