Hello Asier,

you could think about transfecting your cells with mutant huntingtin to induce Drp1-mediated mitochondrial fission in neurons. This paper goes into more detail: http://www.nature.com/nm/journal/v17/n3/full/nm.2313.html

The authors showed that mutant huntingtin binds DRP1 which results in an increased GTPase activity and mitochondrial fragmentation.

With best regards, Christian

Thanks Christian, that is a possibility, but possibly not for my approach. I want to see acute mitochondrial fission during live cell imaging experiments, and check if it is prevented by Drp1 silencing (and therefore if the downregulation is working or not). If I used the mutant htt model I would be very dependant on the transfection efficiency, which is not very high in primary neuronal cultures.

Best,

Asier

What about using Bafilomycin. I have seen in my experiments that Bafilomycin induces mitochondrial fission when used at high concentrations 5 micro molar, although it is an autophagy inhibitor. Acute rotenone treatment also induces mitochondrial fission, depending on the cell type, usually works between 1-10 micro molar concentrations.

Thanks Tanveer! I never tried Bafilomycin nor rotenone, but I will. Rotenone probably will be faster than Bafilomycin right? Another interesting question is if they induce Drp1-denpendent fission (is a mitochondrial fission necessarily Drp1-dependent?).

Best,

Asier