Chavanelle 2015 Fatty Acid Oxidation O2k-Network Discussion Forum

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