Dear all,
We are interested in exploring in vivo mitochondrial functions in disease state. For that we want to track live mitochondria physiological system or in particular tissue or lets say say in blood stream.
We planned to work with mitochondrial protein targeted transgenic mice, but without expertise in this area it seems unclear and difficult to execute.
Does anybody suggest any strategy to do that?
We are also looking for future collaboration on mitochondrial project..
feel free to contact for that..
Thanks all passionate researchers out there..
Cheers,
Raise
Hi, you need to perform mitochondria labeling with pLV-mitoGFP or pLV-mitoRFP (Addgene, LGC Standards Teddington, UK), FACS-sort the cells, amplify, titrate out the signals if you wish, and keep the stock for as long as.
Mitotrack (red)? for mitochondria. It is a kit you may use for mitochondria. I am not sure if it is for in vivo
Hi,
It depends on what physiological parameter you have in mind.
1. You can isolate mitochondria from cells and perform a set of functional assays (i.e. oxygen consumption, ROS production, calcium handling, protein import, etc.) as well as assess structural integrity (i.e. immunoblotting against mitochondrial proteins of interest, mtDNA and/or lipid characteristics etc.).
2. In cells, you can assess some of the mitochondrial parameters including respiration, mitochondrial membrane potential and ROS production using instruments like Seahorse and dyes like TMRM and MitoSOX
3. You can also label mitochondria in vivo (either with mitochondria-targeted fluorescence proteins or with mitochondria specific dies like TMRE) but in this case you are you are likely to assess mitochondrial distribution/morphology in the cells.
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