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Monitoring Metabolic Responses of Single Mitochondria within Poly(dimethylsiloxane) Wells: Study of Their Endogenous Reduced Nicotinamide Adenine Dinucleotide Evolution
journal contribution
posted on 2013-05-21, 00:00 authored by Emmanuel Suraniti, Venkata
Suresh Vajrala, Bertrand Goudeau, Serge P. Bottari, Michel Rigoulet, Anne Devin, Neso Sojic, Stephane ArbaultIt is now demonstrated that mitochondria
individually function
differently because of specific energetic needs in cell compartments
but also because of the genetic heterogeneity within the mitochondrial
pool-network of a cell. Consequently, understanding mitochondrial
functioning at the single organelle level is of high interest for
biomedical research, therefore being a target for analyticians. In
this context, we developed easy-to-build platforms of milli- to microwells
for fluorescence microscopy of single isolated mitochondria. Poly(dimethylsiloxane)
(PDMS) was determined to be an excellent material for mitochondrial
deposition and observation of their NADH content. Because of NADH
autofluorescence, the metabolic status of each mitochondrion was analyzed
following addition of a respiratory substrate (stage 2), ethanol herein,
and a respiratory inhibitor (stage 3), Antimycin A. Mean levels of
mitochondrial NADH were increased by 32% and 62% under stages 2 and
3, respectively. Statistical studies of NADH value distributions evidenced
different types of responses, at least three, to ethanol and Antimycin
A within the mitochondrial population. In addition, we showed that
mitochondrial ability to generate high levels of NADH, that is its
metabolic performance, is not correlated either to the initial energetic
state or to the respective size of each mitochondrion.
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cell compartmentsPDMSMonitoring Metabolic Responsesmitochondrial depositionstages 2mitochondrial populationmitochondrial NADHEndogenous Reduced Nicotinamide Adenine Dinucleotide EvolutionItmitochondrial abilitySingle Mitochondriafluorescence microscopyunderstanding mitochondrialNADH autofluorescenceorganelle levelNADH contentNADH value distributionsStatistical studies
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