If the ROS originate during your Sea Horse Measurement either from the mitochondrial source or extra-mitochondrial sources of the cells, definitely there will be an increase in O2 consumption. This is mainly because of the generation of ROS (superoxide) form molecular Oxygen (O2). In my earlier days, we used to differentiate from the CN-sensitive and CN-insensitive O2 consumption. You should try to block each step of the mitochondrial ET (Complex-I to IV using Cyanide, Oligomycin, FCCP, Rotenone, and Antimycin) and measure O2 consumption and simultaneously the specific ROS (Superoxide and Hydrogen Peroxide) formation. Simply, use SOD-inhibitable Epinephrine (adrenaline) oxidation for superoxide generation in cells under the treatment of different complex-specific inhibitors with cells and under identical conditions, measure oxygen consumption by cells with Sea Horse. Similarly, measure the formation of H2O2 under identical conditions. While measuring the H2O2 production by cells, you may also include a GSH-Peroxidase inhibitor or Catalase inhibitor (azide).

Definitely, ROS production leads to increase in oxygen consumption due to uncoupling of the respiratory ET chain.

Note:

Cyanide is a potent poison that inhibits cellular respiration by binding to cytochrome c oxidase (CCO) in the mitochondria. This inhibition leads to an accumulation of electrons in the electron transport chain, resulting in the production of reactive oxygen species (ROS).

Physiol Rev

. 2014 Jul;94(3):909–950. doi: 10.1152/physrev.00026.2013

Plant Physiol

. 2002 Apr;128(4):1271–1281. doi: 10.1104/pp.010999

There is ample work done in this area.

I just suggested a couple of papers. Please read.

Please read earlier papers published by Dr. Britton Chance and Dr. Hagihara.

I spent decades on this topic. This is my 2 cents worth suggestion.

Good area to investigate, especially in the area of Ferroptosis. Since I am a lipidologist, I am interested in the lipid peroxides and mitochondrial lipids.

Good question (for me).

Parinandi

The Ohio State University Wexner Medical Center

Columbus, OH

Dear Narasimham L Parinandi

Thank you for your detailed and insightful response, and also for your helpful suggestions!