Testing Planaria Neuroregeneration

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msinghal
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Testing Planaria Neuroregeneration

Post by msinghal »

Hello!

This year, I'm researching doing a project testing the effectiveness of different plant compounds in neuroregeneration.
Instead of using human cells, I thought it would be possible to test using planaria.

The different compounds I was considering testing are ashwagandha (Withania somnifera), brahmi (Bacopa monnieri), and lion's mane mushroom (Hericium erinaceus), and the control would be planaria with just food.

I wanted to know how feasible it would be to do this experiment. I was thinking I could measure the effectiveness by how long it takes them to regenerate.

Thanks!
jcschrandt
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Re: Testing Planaria Neuroregeneration

Post by jcschrandt »

Hi, this is a very interesting project! Planaria would be a good model for studying regeneration, but how are you planning to observe regeneration of the nervous system? When I've done studies with C. elegans, we usually use fluorescent tags to visualize a protein or cell-type. Do you have access to a fluorescent microscope? Other methods for labeling the nervous tissue involve staining, but they would kill the flatworm.

If you just want to observe how long it takes to regenerate when fed these different compounds, then you would be studying how the compounds affect stem cell division and differentiation. This would also be a very interesting study!

I hope this helps!
Jen
msinghal
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Joined: Fri Jun 21, 2013 9:37 am
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Re: Testing Planaria Neuroregeneration

Post by msinghal »

Dear Jen,

I'm going to ask our biotech teacher on the 19th if there's a fluorescent microscope.
Since it's too close to the competition date to contact any local labs, I was thinking the easiest way to test regeneration was by how fast the planaria grew. In this case, I would still only want to be testing one variable at a time. Since doing this would be measuring neuroregeneration as well as stem cell behavior, could I just broaden the question of the experiment or would it have to be more specific?

Thanks!
PharmaMan
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Re: Testing Planaria Neuroregeneration

Post by PharmaMan »

Hello,

This would still be an interesting topic to investigate using the model system you have described with the different compounds. However, unless you have a way to identify different components of the nervous system as Jen described, you wouldn't be able to make any conclusions about regeneration of neurons.

For the focus of your project, I wouldn't keep it too broad. The ultimate goal will be to answer a specific research question with sufficient rigor; therefore, you will need a specific question to investigate using a limited number of variables that will allow you to either support or refute your hypothesis.

What are you more interested in testing at this point? Let us know and we'll be happy to help further.

Best,
Pharma
msinghal
Posts: 15
Joined: Fri Jun 21, 2013 9:37 am
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Project Question: Microbial Fuel Cells
Project Due Date: Spring 2014
Project Status: I am just starting

Re: Testing Planaria Neuroregeneration

Post by msinghal »

Hello,

I still want to study the effects of the plants on only neural regeneration; if I used something like this: http://www.carolina.com/platyhelminthes ... /132912.pr
would I be able to identify the nervous system with a microscope? In my experiment, I wanted the independent variable to be which compound the planaria are exposed to, with a control of nothing added.

Thanks!
PharmaMan
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Re: Testing Planaria Neuroregeneration

Post by PharmaMan »

Hello,

I am not sure if this would be a good system to use. I looked at the website; however, I am hesitant because it does not say what the intention of the pre-staining is supposed to do. I feel a better experiment would probably be to test cultured neurons instead to completely isolate these cells from other cell types/tissues within the model organism. Do you have a way that you may be able to carry out an experiment like this?

Let us know and we'll be happy to help you further. Best of luck!
Pharma
msinghal
Posts: 15
Joined: Fri Jun 21, 2013 9:37 am
Occupation: Student, 8th Grade
Project Question: Microbial Fuel Cells
Project Due Date: Spring 2014
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Re: Testing Planaria Neuroregeneration

Post by msinghal »

Hello,

The only nearby place I can think of where I can experiment with actual neurons is my local community college, and they only let registered students access the labs.

I was thinking, since these plants are mainly used enhance memory etc, instead of using planaria specifically to test neural regeneration, would it make sense to test the effects of the plants on the planaria's ability to perform some cognitive task?
I could test how fast it takes the planaria to navigate a maze (or something along those lines) before and after (with a different maze) the plant extract.

Would testing the above idea or the plant's effects on general planarian regeneration be more scientifically rigorous?

Thanks!
PharmaMan
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Re: Testing Planaria Neuroregeneration

Post by PharmaMan »

Hello,

You could test these extracts on planaria; however, I am not sure if carrying out a cognitive experiment with planaria would work. Do you know what some of the other functions of these extracts are? If they have the ability to stimulate re-growth or growth in general on the worms, I would think that this may yield better results for your experiment.

Let us know what you plan to do and if you have further questions.

Best,
Pharma
msinghal
Posts: 15
Joined: Fri Jun 21, 2013 9:37 am
Occupation: Student, 8th Grade
Project Question: Microbial Fuel Cells
Project Due Date: Spring 2014
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Re: Testing Planaria Neuroregeneration

Post by msinghal »

Hello,

I think these compouds only affect neural regeneration by through interacting with the production of neurotransmitters.
In this experiment, they tested the negative phototactic response of planaria during different stages of head regeneration: http://www.bioone.org/doi/pdf/10.2108/zsj.21.275

I was thinking by testing the planaria's response to light I would be indirectly testing the progress of the head regeneration.

Thanks!
PharmaMan
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Re: Testing Planaria Neuroregeneration

Post by PharmaMan »

What types of planaria are you going to be using for this study? If these compounds are supposed to support regeneration of neurons, you would need a model system that is deficient in some respect with nervous system development/growth. If these compounds are supposed to deteriorate neuron function, then a wild-type planaria model would work.

In terms of testing their response to light, you would be testing a very specific portion of the nervous system. To test the regeneration of neurons involved in processing visual stimuli, you would have to find a way to isolate this part of the nervous system using molecular biology techniques. However, you could use their response to light as an indirect measure of head regeneration. I would be cautious in the sense that head regeneration will be a process that involves much more than just the nervous system, and these compounds that you are testing may have no effect/detrimental effects on these other germ layers involved in the regeneration process.

Let us know if you have further questions. Good luck!
Pharma
jcschrandt
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Re: Testing Planaria Neuroregeneration

Post by jcschrandt »

Hi, I think that the negative controls and molecular biology studies would be good next steps, but I don't think they're necessary for your initial experiment. I think you have a good idea about treating planaria with different compounds to see how they react to light. Here are a few suggestions:
1. Follow the phototaxis procedure in the article you attached as closely as possible. Since you probably can't autoclave water, you may be able to boil purified water to sterilize it.
2. Carefully consider how you are going to treat planaria with the compounds. Are they going to eat the compounds? Are these compounds things that planaria would naturally eat? Would the compounds provide the right nutrients for the planaria?
3. Any differences in phototaxis may be very small, so you should consider how frequently you will test them. You may want to test them more frequently than the authors of the article.
4. As PharmaMan mentioned, take into consideration that the compounds may affect more than just neurons and that head regrowth affects more than just neurons.

Best regards,
Jen
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