I want to treat my worms with taxol by growing them on NGM plates containing it and then test their sensitivity to light touch, in the converse of the colchicine experiment, but I can’t seem to find any references that describe doing this. Has anyone tried? Does it even get through the cuticle at any concentration? Any idea what concentration of taxol might affect the TRNs but not cause gross abnormalities?
I’d thought I recalled people trying it before, but all I could find was this paper, which reports that even the maximum dose they tried (1 uM) didn’t seriously inconvenience the animals (they report statistically significant effects, but they don’t appear to be especially striking).
Oh well. Thanks Hillel.
This paper (http://www.molbiolcell.org/content/early/2012/12/03/mbc.E12-06-0441.long) also reports the use of taxol to stabilise microtubles in TRNs.
The authors use a [i]bus-17[i] mutant background to enhance the permeability of the drug through the cuticle.
The protocol is pretty standard, the drug is just added to regular NGM.
Thanks. Yes, I had seen that paper, but I can’t find the concentration for the paclitaxel. They reference a Chalfie 1982 paper for the method, but that paper doesn’t have any paclitaxel treatments. Puzzling.
yes you’re correct, there doesn’t appear to be a concentration mentioned anywhere and the reference to the Chalfie paper seems just with a view to referencing the ‘general approach’ e.g. using colchicine and similar plates in assays à la Chalfie et al.!
I remember having seen the concentration for paclitaxel mentioned in other papers (50µM??), but these were working with embryos not worms.
Perhaps it would be best to email the authors of the paper suggested by wrm-99 and ask them what concentration they used? Then let us all know!
I just wanted to follow up and say that I did email the authors, and they immediately replied that the concentration used was 500 nM (dissolved in DMSO before adding it to the liquid NGM), apologized for accidentally deleting it from the final copy, and kindly added a few extra pointers regarding how to do the experiment.
Great…thanks for finding that out and posting it on the forum…now we all have a quick reference should we ever want to use taxol.