We have a transgenic KO mouse model for ALS and are wanting to look at mitochondria as a possible indicator of retrograde transport disruption in motor neurons. We have longitudionally sectioned muscle tissue to use for the preliminary work that we normally use to look at the neuromuscular junction. All of the literature I've been able to find is either about mitochondria in the neurons, or in the muscle but nothing about the junction.
Any suggestions would be greatly appreciated.
If tissue is fixed, some MitoTracker dyes will label mitochondria, or - label with anti-OxPhos Antibodies [see link below]. You will need confocal imaging and some other marker for the motor nerve terminal, so that you can distinguish between mitos IN the terminal and other labeled mitos in muscle, Schwann cells ... For labeling mitos in live motor nerve terminals there are more possibilities..
http://tools.lifetechnologies.com/content/sfs/manuals/mp07510.pdf
Thank you for the suggestions. Unfortunately we do not have an EM. Gavriel, we took your advice and are using MitoTracker and labeling the nerve terminal. So far it is going smoothly! Much appreciated!
May be too late, but hope these tips will help someone who have the same question:
NMJ is not easy to get when you section the muscle. What I learn from a friend is that you can take a 1-5 mm (NOT um !!!) thick muscle and clip it between 2 glass for 2 hours (up to 1 day) in a wet box to keep moisture. penetrate with Triton x-100 O/N and proceed for immunostaining. NMJ can be labled with FL-conjugated BTX easily in this way.
For mitochondria colocalization with BTX, you'd better use a neuronal specific GFP-mito mouse line such as mito-dendrite mouse and staining with GFP antibody. Or you can use an axonal specific mitochondria marker like syntaphilin to do the mitochondria co-staining.
http://www.jbc.org/content/286/26/23432.abstract
http://www.cell.com/neuron/abstract/S0896-6273%2815%2900569-3
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