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Talk:Fatty acid oxidation

From Bioblast

O2k-Network discussion forum: fatty acids used in permeabilized fibre assays (2015-10-20)









FA and cell toxicity

I am working at the University of Colorado with Drs Paul MacLean and Matthew Jackman who acquired two OROBOROS this year. Following our collaborators advice (Dr Donal O'Gorman at Dublin City Uni), we are following your protocol to measure fat oxidation. Contrary to results in the literature and from Dr O'Gorman's lab, as soon as we add L-palmitoylcarnitine, the cells die. 2 uM only of L-PC are sufficient to reach a respiration of zero. Adding any other substrates does not allow an increase in O2 consumption. Our local collaborator Dr Jane Reush is encountering the same issue and therefore only use octanoyl carnitine. Octanoyl carnitine does not alter mitochondria/cell. Following advices from Dr Neufer (North Carolina), we added some carnitine. This did not allow maintaining the integrity of the cell. (1)

Because we are in altitude (1600m), we need to use a piO2 of 200-220 (instead of 160) in basal and of 350 (instead of 300) in maximal. We are wondering if this higher piO2 could promote the generation of ROS species or oxidative stress that would kill the mitochondria/cell when adding L-PC? (2)

Audrey (2015-05-14)
O2k-Network: US CO Aurora Jackman MR
Audrey Bergouignan, PhD
Anschutz Health and Wellness Center Division of Endocrinology, Metabolism and Diabetes
University of Colorado
E [email protected]
http://www.anschutzwellness.com/​


Response

  1. Β»Fatty acid oxidation#FAO and HRR
  2. When using permeabilized muscle fibers, we have to increase oxygen levels to nonphysiological hyperoxia to avoid oxygen diffusion limitation to the fibers. Although high oxygen levels stimulate ROS production, mitochondria remain functional during times of measurement up to 3 hours. (Gnaiger E, 2015-05-15)