Larsen 2017 MiP2017 WG2

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Steen Larsen
The effect on respiratory capacity using different respiration buffers.

Link: MiP2017

Larsen S (2017)

Event: MiP2017

COST Action MITOEAGLE
Mitochondrial respiratory capacity and the sensitivity for ADP was measured in permeabilized human skeletal muscle fibers using MiR05 with or without blebbistatin before and after 6 weeks of high intensity training (n=12; 7x1 min at 100% of VO2max – 3 times a week). Furthermore mitochondrial respiratory capacity was measured in permeabilized human skeletal muscle in either buffer Z or MiR05 (n=8).

The muscle biopsy was obtained using the Bergstrom technique. Immediately after the muscle was placed in BIOPS, and was within 20 min dissected using sharp needles, followed by saponin permeabilization (30 min) on ice, finally the fiber bundles was washed 2 x 10 min in either MiR05 or buffer Z (on ice) before weighed and placed in the O2k-FluoRespirometer (Oroboros Instruments).

Mitochondrial respiratory capacity was lower in MiR05 when blebbistatin was not present (both pre and post training) with complex I and II linked substrates (mal-glut-ADP-succ). Apparent Km for ADP was higher in the present of blebbistatin pre training. No changes was seen with training when blebbistatin was present, whereas Km increased with training when blebbistatin was not present.

When comparing mitochondrial respiratory capacity in different buffers (buffer Z and MiR05), some differences occurred. Mitochondrial respiratory capacity with complex I linked substrates were higher in buffer Z compared with MiR05, whereas no differences were seen when complex I + II linked substrates were analysed together.


Bioblast editor: Kandolf G O2k-Network Lab: DK Copenhagen Dela F, DK Copenhagen Larsen S


Labels: MiParea: Respiration, Exercise physiology;nutrition;life style 


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


Coupling state: OXPHOS  Pathway: N, NS  HRR: Oxygraph-2k 


Affiliations

Xlab, Center Healthy Aging, Dept Biomedical Sciences, Fac Health Sciences, Univ Copenhagen, Denmark. - stelar@sund.ku.dk

Figure 1

LarsenS figure MiP2017.jpg