I'm running experiments and I need to silence the mouse DRG in culture. I'm thinking of either Lidocaine or Toxin cocktail (TTX with conotoxins). Does any one have experience in this and willing to share some information regrading protocol conc. duration etc.
Hi Rania,
Topical lignocaine or lidocaine, or equivalent local anestehetic, application sounds like the most reasonable idea, cheaper and possibly more effective. Local application anywhere around the DRG or between the DRG and dorsal horn of the spinal cord would be a perfect location. As you know these local anesthetics will block the Na channels and action potential conduction along the axon and therefore you do not need to worry about the TTX-resistant Nav subunits expressed in the soma. How specific are you trying to be? Just a single segment/DRG or multiple?
Sorry I just realized that you are doing it on cultured neurons. Why not in vivo? I would just use TTX in that case around 1-10 micromolar should block action potentials. See my 2005 JPET electrophysiology paper on cultured DRG neurons. Another drug you may consider is riluzole, an FDA approved drug, though it does not block all action potentials (see my 2013 NeuroMethods paper).
Hi Refik,
Thank you for your replay and suggestion. what about TTX-R in DRG culture?? do you have idea to block them? and what is your opinion about using lidocaine to block DRG in culture?
HI Rania,
That depends on what question(s) you are asking in your experiment. Are trying to block all Nav channels or just block action potential firing or activity. I am aware of TTX-R are in some DRG neurons, but are you sure that your cells are expressing TTX-R in culture? I would just try TTX first and if you do not get the result than try conotoxins etc which may be a bit expensive to use.
best wishes, Refik
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