title: Monitoring metabolic responses of single mitochondria within poly(dimethylsiloxane) wells: study of their endogenous reduced nicotinamide adenine dinucleotide evolution.
abstract: 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.
authors.affiliation: University Bordeaux, ISM, UMR5255, F-33400 Talence, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Suraniti
authors.forename: Venkata Suresh
authors.initials: VS
authors.lastname: Vajrala
authors.forename: Bertrand
authors.initials: B
authors.lastname: Goudeau
authors.forename: Serge P
authors.initials: SP
authors.lastname: Bottari
authors.forename: Michel
authors.initials: M
authors.lastname: Rigoulet
authors.forename: Anne
authors.initials: A
authors.lastname: Devin
authors.forename: Neso
authors.initials: N
authors.lastname: Sojic
authors.forename: Stephane
authors.initials: S
authors.lastname: Arbault
mesh_terms: D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D046228:Microarray Analysis / Q000379:methods*; D008928:Mitochondria / Q000378:metabolism*; D009243:NAD / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology; D013050:Spectrometry, Fluorescence
chemical_list: D004129:Dimethylpolysiloxanes; D009243:NAD; C013830:baysilon