Hello, Now, I’m doing experiments about the effect of drug candidate. I want to see the effect of drug to only microglia, not neurons. If I do inject drugs to mouse groups, it may down regulate microglial activation and at result, protect dopaminergic neurons from MPTP(toxic chemical) in Substantia nigra region. But these method cannot exclude direct effect of drug to neurons. How to exclude direct drug effect to neurons? Please help me!:(
First, I commend you on doing this in vivo experiment as it is very relevant. In vivo experiments are inherently to observe if the effect on the desired outcome is evident in animal/organismal model with or without measurable side effects. Your outcome here is neuronal viability/morphology/intactness/markers not affected or less severely affected by MPTP. This could be due to your hypothesis of suppression microglial activation, but numerous other factors may also be involved including the one you suggested on direct effect on neuronal cells.
I am afraid that you will not be able to tell this distinction from the experimental model you suggested but can do this effectively in vitro. You can expose your neurons to MPTP and see if your drug can alter the toxicity profile. You can do similar experiments on microglial cultures and may even try co-cultures. A mechanism of action established in vitro will help you to explain evens in vivo.
Solomon Habtemariam
Thank you for your advice so much.
I think it may be explained in vitro as you mentioned and will use co-culture system with neurons.
THANK YOU FOR YOUR HELP!