Monitoring
Metabolic Responses of Single Mitochondria
within Poly(dimethylsiloxane) Wells: Study of Their Endogenous Reduced
Nicotinamide Adenine Dinucleotide Evolution
Emmanuel Suraniti
Venkata
Suresh Vajrala
Bertrand Goudeau
Serge P. Bottari
Michel Rigoulet
Anne Devin
Neso Sojic
Stephane Arbault
10.1021/ac400494e.s001
https://acs.figshare.com/articles/journal_contribution/Monitoring_Metabolic_Responses_of_Single_Mitochondria_within_Poly_dimethylsiloxane_Wells_Study_of_Their_Endogenous_Reduced_Nicotinamide_Adenine_Dinucleotide_Evolution/2412817
It 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.
2013-05-21 00:00:00
cell compartments
PDMS
Monitoring Metabolic Responses
mitochondrial deposition
stages 2
mitochondrial population
mitochondrial NADH
Endogenous Reduced Nicotinamide Adenine Dinucleotide EvolutionIt
mitochondrial ability
Single Mitochondria
fluorescence microscopy
understanding mitochondrial
NADH autofluorescence
organelle level
NADH content
NADH value distributions
Statistical studies