Chavanelle 2015 Fatty Acid Oxidation O2k-Network Discussion Forum

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Chavanelle 2015 Fatty Acid Oxidation O2k-Network Discussion Forum

Link: O2k-Network Discussion Forum

Chavanelle V (2015)

Event: Fatty Acid Oxidation O2k-Network Discussion Forum

Fatty oxidation in permeabilised db/db mice muscle fibres, in Mir05 + Cr +37°C. We found that ADP-stimulated respiration in presence of 0,02 mM of palmitoyl-carnitine (OXPHOS) is very close to the non-ADP-stimulated respiration (LEAK state) when working with these conditions. (Values around 14 to 20 (pmol/s/mg w.t.) for LEAK state and 15 to 24 (pmol/s/mg w.t.) for fatty acid triggered-OXPHOS). These data suggest either a very poor ability for this animal model to use palmitoylcarnitine or a very high LEAK state (or both!)

Protocol

  1. 2,2 mL mir05+Cr was added in the chambers and oxygen was added to the medium up to 400 – 450 nmol/mL
  2. Chambers were closed
  3. 10µM Amplex Red was added to the chambers
  4. 2 U HRP was added
  5. 10 U SOD was added
  6. 20µM Palmitoyl-Carnitine was added
  7. 2 mM Malate was added
  8. 1 to 3 mg wet permeabilised muscle fibers was added
  9. LEAK state was recorded
  10. Mg+ was added
  11. 2,5 mM ADP was added
  12. Pal-C stimulated OXPHOS was recorded

The tissue used was skeletal muscle (Gastrocnemius)

Keywords: Amplex Red


Labels: Pathology: Diabetes 

Organism: Mouse  Tissue;cell: Skeletal muscle  Preparation: Permeabilized tissue 


Coupling state: LEAK, OXPHOS  Pathway: F, N 


Palmitoylcarnitine