Introducing the NextGen-O2k Oroboros with a focus on coenzyme Q

Abstract

Mitochondria are recognized as essential players in virtually every physio-pathological context.

We here present Oroboros NextGen-O2k: the complete experimental system for high-resolution fluorespirometry and redox biology enabling simultaneous monitoring of oxygen consumption together with coenzyme Q redox state, NAD(P)H redox state, H2O2 production (AmR), mt-membrane potential (TMRM, Safranin and Rhodamine 123), Ca2+ uptake capacity (CaG), ATP production (MgG) or photobiology measurements. Substrate-uncoupler-inhibitor titration protocols have been developed to study the redox states of the mitochondrial coenzyme Q pool. A three-electrode sensor has been implemented into the NextGen-O2k to monitor continuously the redox state of CoQ2 added as a Q-mimetic simultaneously with O2 consumption.

The Q-Module was optimized for high signal-to-noise ratio, minimum drift, and minimum oxygen diffusion. CoQ2 equilibrates in the same manner as endogenous CoQ at Complexes CI, CII and CIII. The CoQ2 redox state is monitored amperometrically with the working electrode, which is poised at CoQ2 redox peak potential determined with cyclic voltammetry. The voltammogram also provides quality control of the Q-sensor and reveals chemical interferences.

Find more info at https://www.oroboros.at/

About the speaker

Luiza Cardoso is a PI in the scientific team of the company Oroboros Instruments.

She completed her Master’s and PhD studies at the Biophysics Institute of the Federal University of Rio de Janeiro, Brazil, studying P-type ATPases and regulation by protein kinases, and copper homeostasis. She moved to Austria in 2018 for a PostDoc, first at the Medical University of Innsbruck then at Oroboros Instruments. There, she has been involved in different research projects studying mitochondrial physiology and in the development of protocols for analysis of ATP production with the fluorescent dye Magnesium Green, and of coenzyme Q and NADH redox biology together with respirometry with the NextGen-O2k, and the SUITbrowser – a platform designed to help researchers find a protocol for respirometry.

She is currently co-editor of the journal Bioenergetics Communications and MitoFit Preprints.