Mitochondrial stress induction protocols.
What about other protocols for oxidative stress induction by using H2O2?
You can induce oxidative stress by number of methods, but each of them generally have side effects. For instance, ethidium bromide causes wide range of effects (including mutations). I believe that use of specific mitochondrial electron transport inhibitors (at appropriate concentrations) can promote generation of ROS by mitochondria. However, if you use these inhibitors at higher concentrations, they can have multiple effects. You may consult attached reviews (that impressed me a lot) on mitochondria.
Are you trying to induce mitochondrial stress, deplete mitochondria, or a combination of both? Some stressors may activate mitochondrial biogenesis and result in more mitochondria. What is the objective of the experiments you want to perform?
I am trying to induce stress in the cells by targeting mitochondria.
by the way can you tell me about those stressors that may activate mitochondrial biogenesis?
There are a number of pharmacological agents that will stress the mitochondria by inhibition at the electron transport chain subunits such as rotenone (complex I inhibition), 3-nitropropionic acid (complex II), antimycin (complex III), cyanide (complex IV) or oligomycin (ATP synthase inhibitor).
There are a few papers showing mitochondrial biogenesis following oxygen glucose deprivation in brain cells and also following ischemic injury in rodent cerebral stroke models.
Dear Prof. Jordan Kolarov- can you please let me know any published paper or scientific record stating that EtBr can specifically deplete mtDNA. Do you really believe that EtBr will not influence nuclear DNA and DNA present in plastids? What do you think is the mechanism by which EtBr depletes DNA or mtDNA? Please suggest specific references.
If you put in Google (rho zero cells), you will get a number of relevant publications on both mammalian and yeast cells answering your questions. Here is just one example -Methods Mol Biol. 2009;554:383-91. doi: 10.1007/978-1-59745-521-3_23.
Establishment of human cell lines lacking mitochondrial DNA.
Hashiguchi K1, Zhang-Akiyama QM.
Another question I concern is whether EtBr can also deplete nuclear DNA as well? For instance I want to deplete my mtDNA but preserve nuclear DNA? Any ideas?
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