Hi,
I classically performed tissue sampling in vivo to isolate mitochondria from skeletal muscle. However, the reviewer of the ethic committee doesn't want that we performe the tissue sampling on living animal. Does anybody have some articles to demonstrate the interest of doing tissu sampling on living animal.
Many thanks
If you take a muscle biopsy for mitochondrial isolation, a clear advantage is that the animal stays alive. You can also isolate mitochondria from the muscle of a euthanased animal (provided you are using a method of euthanasia that does not interfere with mitochondrial function and collect the tissue quickly). From the viewpoint of mitochondria there is no much difference, but from the viewpoint of the animal, there clearly is one.
Dear Inna M Sokolova,
Could you provide more details about the euthanasia procedures that doesn't interfere with mitochondrial function ?
I know isoflurane is usually recommanded but I'm wondering for instance if isoflurane overdose is ok to euthanise mice before mitochondrial isolation ?
Hi Frederic - big fan of your work!
We have done all of our animal work on tissue collected post mortem, and as long as the tissue is collected quickly after death, we dont see any problems. This is with a variety of different methods of euthanasia - some anaesthetics, like isoflurane or pentobarbitone work by uncoupling mitochondria, but these effects “wear off” when you prepare the tissue. In mice/rats, I haven’t noticed a difference between isoflurane, pentobarbitone, and cervical dislocation.
Biopsy is good if you want to compare before and after an intervention in the same animal, but is quite hard in rodents - easier in people!
Mitochondria can be easily isolated from skeletal muscle post mortem. It’s absolutely no problem with respect to mitochondrial integrity and function.
There is no reason to collect tissues from living animals.
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