Hi folks
I am trying to study the effect of exercise on Parkinson's disease rat model. For this reason, I am trying to look at neuron from the cortex and the striatum to do some western and RT-PCR. The problem is that I couldn't find a single paper talking about doing so on adult rat (e.g. 3 or 6 months old). Because there is no point of growing neuron from E13-E17.
The second question is about sorting neuron since neuron will be mixed with glial cells, do you think that sorting them with flow cytometry is doable?
Thanks in advance
Yes,but in our experience they all die within 14 days.
Use the chemically defined medium and glia growth is seriously reduced, or use ArabC. Good luck
Hello Mohannad,
I found it challenging to culture adult CNS neurons but it is definitely possible. This protocol is useful: Article Brewer, G.J. & Torricelli, J.R. Isolation and culture of adu...
I think it is extremely difficult to culture adult CNS neurons after sorting them through flow cytometer. In my experience, they usually don't survive after flow cytometry and don't grow well (/at all). The above protocol uses Optiprep density gradient media that allows for separation of neurons from non-neuronal cells, so that might help.
Good luck!
Everything in possible and what you intend is possible as indicated by Enrico and Shrinivasan, but for me you have to decide on doing research on i) how to improve the method for primary cultures of neurons from adult aimals or ii) research on Parkinson. In other words, almost every paper on primary cultures of neurons for Parkinson's or Alzheimer's research are obtained from fetuses.
Dr. Marani
Thank you for your response, if I can keep them for 14 days that means I need to sacrifice a lot of rats.
Mr. Raghuraman,
the reference you posted was the one I am planning to follow and the first thing that came up to my side was using the flow cytometer to get rid of glial cells. Dr. Marani mentioned a nice point of using a chemically defined medium to reduce glial cells growth and I think I will go with that to reduce the loss of the neuron.
Dr. Franco,
First of all thank you so much for the points you brought up. I am planning to look at how neuron from exercised rat would respond to stress and mitochondria toxins, for that reason I was looking for any paper used primary neuron from adult rat (6 months old).
Thank you all for your advices
By the research you are conducting one option (to me) is to look for expression of significant proteins of oxidative stress and of innate antioxidant mechanisms by western blotting using samples from brain. Best wishes Mohannad
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