I would like to study mitochondrial dysfunction in MutUNG1 mice ,GPR81 KO mice which approach would be better? or generally to study the mitochondrial dysfunction, which suggestions would you have?
What type(s) of mitochondrial dysfunction are you interested in? Mitochondria do many things, so you could test mitochondrial dysfunction in a variety of ways - metabolic perturbations (including but not limited to oxygen consumption rates, ATP levels, etc), ROS generation, intrinsic apoptotic induction, differences in morphology, maintenance of mtDNA, and so on. Do you have a specific question you are trying to answer or hypothesis you are trying to test? Are you limited to mouse tissues/in vivo models or do you have cell lines to work with?
What type(s) of mitochondrial dysfunction are you interested in? Mitochondria do many things, so you could test mitochondrial dysfunction in a variety of ways - metabolic perturbations (including but not limited to oxygen consumption rates, ATP levels, etc), ROS generation, intrinsic apoptotic induction, differences in morphology, maintenance of mtDNA, and so on. Do you have a specific question you are trying to answer or hypothesis you are trying to test? Are you limited to mouse tissues/in vivo models or do you have cell lines to work with?
The easiest way is to do morphological study. This can be done by fluorescence microscopy. You can use genetically encoded proteins or simple fluorescent markers for this purpose. It really depends what type of information you want to get. There are ways to study many aspects (mentioned by Natalie) of mitochondria using several different approaches.
In mouse model the easiest tissue to study mitochondria is liver. You can obtain mitochondria from muscle, heart and brain too, but you will need to practice until obtain a good RC to your mitochondria. One of the most simple assay to study mitochondrial swelling challenged with calcium. This isn't so specific, but you can associate this method with those that Natalie suggested.
Many Methods are available
1. Measure the oxygen consumption by using Mitocell (clark oxygen electode).
2. Monitor mitochodria shape and arrangement by using Mitotracker dyes to the tissues.
3. You can take cardiac muscle for better understanding.
so many methods are available from outer to inner and the dysfunction of mitochondria causes generally and finally and not limited of course in apoptosis.
all enzymes and process near or linked to mitochondria are estimated,,, and so for (cell physiology, cellular death, apoptosis, caspases....etc.) are the approaches to study.
Dear Dr. David Aphkhazava,
One of the strategies could be to check mitochondrial associated Ca2+ signaling using microscopic methods of Ca2+ yield by FRAP.
mtDNA content could also be checked.
We have done some work on mitochondrial dysfunction which can be found in the following articles:
Hope this helps you out
Data Role and clinical significance of lymphocyte mitochondrial d...
Article Mitochondrial Oxidative Stress-Induced Epigenetic Modificati...
You can study the mitochondrial membrane potential using DIOC6 dye.
It's easy to carry out after cell culture.
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