I am testing the effects of a drug, soluble in DMSO, on electrophysiological properties of dopamine neurons. The experimental protocol consists of 110 min of incubation. Since this drug is only soluble in DMSO, I first check whether the sole incubation with DMSO for 110 min may causes changes of neuronal intrinsic properties. Following this incubation, my first impression is that neuronal membranes are more fragile; it is harder to obtain a cell-attached recording, and the membrane easily goes to rupture after gaining the seal.
Does anyone have experience with this kind of issue? Are there any useful tips to handle a prolonged incubation with a drug soluble in DMSO?
DMSO can seriously damage membranes and I think it could contribute to what you experienced. Are you using > 0.1% DMSO in the final solution? Some of DMSO soluble drugs can be dissolved in EtOH. Have you tried that?
When I used 0.05% DMSO neurons were perfectly fine for 40min. It seemed to me that 0.1% DMSO at 35°C changed quality of the membrane although the neurons survived. It can be specific for the neuronal type and the incubation temperature. Of course, use the same DMSO concentration (withouth the drug) in a control recording. It is also good to consider, if the drug stays soluble in low DMSO concentration. Good luck.
I agree with all comments. We tried to use long incubation times (> 40 min) with DMSO 0.1 % but neurons got leaky and unstable. Try ethanol instead if possible.
Regards
Thank you very much to all of you for the suggestions.
Yes, I'm using DMSO 0.1%. For the same project I'm testing several compounds only soluble in DMSO, that's why I haven't tried the EtOH yet.
I'll do it at least for the one is soluble in EtOH. Do you know any good inhibitor of endocytosis that I can use intrapipette for patch clamp-recordings?
Thanks a lot once again!
Hello,
You may try this :
http://www.sigmaaldrich.com/catalog/product/sigma/d7693?lang=fr®ion=FR
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