Simon
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Here's a bit more about the Seahorse technology and what it measures, using info from the Seahorse site. First off, it's looking at the body's two main energy pathways, glycolysis and respiration, which you can see in these diagrams (from Seahorse videos).
First 55 seconds of this video is good on the underlying biology
But this micro-chamber is only a tiny amount of the fluid in each compartment. After each measurement, the probes are raised, everything mixes together and it can then take a new measurement. This ability to take repeated measures over time is key.
This study measures a bunch of different aspects of oxidative phosphorylation, which it does by injecting different compounds to probe what's really going on. Think of it as an exercise test for mitochondria, revealing what it's doing and how much capacity it has to ramp up activity. The central measures in the study are:
1. Basal respiration, the tickover rate of activity
2. ATP production, which is simply basal respiration less a bit of inefficiency loss (proton leak)
3. Maximal respiration: what the mitochondria can theoretically do using a drug to put the foot to the floor.
4. Reserve capacity: simply maximal respiration less basal respiration (peak capacity less what it's doing, or headroom to increase activity).
Fig 1 from the paper shows an example of how this testing works but I think my version is a bit simpler (the names at the top are the compounds added to put mitochondria through their paces).
The study looked at glycolysis too, which uses a slightly different detailed methodology, but that found nothing of interest, so I will spare you the details.
First 55 seconds of this video is good on the underlying biology
- Glycolysis produces modest amounts of ATP, the cells' main fuel as well as pyruvate, the molecule that feeds mitochondria. Glycolysis produces a lot of protons (acid, effectively, often as lactic acid).
- Oxidative phosphorylation (or respiration) is the process that takes place in mitochondria, the real fuel factories that burn fuel efficiently, consuming oxygen and foodstuffs and pumping out ATP.
But this micro-chamber is only a tiny amount of the fluid in each compartment. After each measurement, the probes are raised, everything mixes together and it can then take a new measurement. This ability to take repeated measures over time is key.
This study measures a bunch of different aspects of oxidative phosphorylation, which it does by injecting different compounds to probe what's really going on. Think of it as an exercise test for mitochondria, revealing what it's doing and how much capacity it has to ramp up activity. The central measures in the study are:
1. Basal respiration, the tickover rate of activity
2. ATP production, which is simply basal respiration less a bit of inefficiency loss (proton leak)
3. Maximal respiration: what the mitochondria can theoretically do using a drug to put the foot to the floor.
4. Reserve capacity: simply maximal respiration less basal respiration (peak capacity less what it's doing, or headroom to increase activity).
Fig 1 from the paper shows an example of how this testing works but I think my version is a bit simpler (the names at the top are the compounds added to put mitochondria through their paces).
The study looked at glycolysis too, which uses a slightly different detailed methodology, but that found nothing of interest, so I will spare you the details.
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