JoVE   
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Biology

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Neuroscience

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Immunology and Infection

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Clinical and Translational Medicine

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Bioengineering

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Applied Physics

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Chemistry

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Behavior

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Environment

|   

JoVE Science Education

General Laboratory Techniques

You do not have subscription access to videos in this collection. Learn more about access.

Basic Methods in Cellular and Molecular Biology

You do not have subscription access to videos in this collection. Learn more about access.

Model Organisms I

You do not have subscription access to videos in this collection. Learn more about access.

Model Organisms II

You do not have subscription access to videos in this collection. Learn more about access.

Essentials of
Neuroscience

You do not have subscription access to videos in this collection. Learn more about access.

Essentials of Developmental Biology

You have subscription access to videos in this collection through your user account.

 JoVE Biology

Micro-dissection of Rat Brain for RNA or Protein Extraction from Specific Brain Region

1, 1, 1, 1, 1

1Laboratory of Neurodegenerative Diseases, Department of Anatomy, LKS Faculty of Medicine, The University of Hong Kong - HKU

Article
    Downloads Comments Metrics

    You must be subscribed to JoVE to access this content.

    This article is a part of   JoVE Biology. If you think this article would be useful for your research, please recommend JoVE to your institution's librarian.

    Recommend JoVE to Your Librarian

    Current Access Through Your IP Address

    You do not have access to any JoVE content through your current IP address.

    IP: 184.73.104.82, User IP: 184.73.104.82, User IP Hex: 3091818578

    Current Access Through Your Registered Email Address

    You aren't signed into JoVE. If your institution subscribes to JoVE, please or create an account with your institutional email address to access this content.

     

    Summary

    Micro-dissection of rat brain into various regions is extremely important for the study of different neurodegenerative diseases. This video demonstrates micro-dissection of four major brain regions include olfactory bulb, frontal cortex, striatum and hippocampus in fresh rat brain tissue. Useful tips for quick removal of respective regions to avoid RNA and protein degradation of the tissue are given.

    Date Published: 8/30/2007, Issue 7; doi: 10.3791/269

    Cite this Article

    Chiu, K., Lau, W. M., Lau, H. T., So, K., Chang, R. C. Micro-dissection of Rat Brain for RNA or Protein Extraction from Specific Brain Region. J. Vis. Exp. (7), e269, doi:10.3791/269 (2007).

    Abstract

    Micro-dissection of rat brain into various regions is extremely important for the study of different neurodegenerative diseases. This video demonstrates micro-dissection of four major brain regions include olfactory bulb, frontal cortex, striatum and hippocampus in fresh rat brain tissue. Useful tips for quick removal of respective regions to avoid RNA and protein degradation of the tissue are given.

    Protocol

    Step 1

    Sacrifice the rat according to established protocols. Remove the brain from the skull and rinse it in ice cold DEPC treated Milli Q water to remove any surface blood.

    Step 2

    Place on cold metal plate. Cut the brain bi-half into right and left hemisphere.

    269_Figure-01

    Figure 1.  The cutting positions from the medial view of rat hemisphere
     

    Step 3

    Collect the olfactory bulb right after the first cut. Flash freeze the specimen in liquid nitrogen and store at -80°C.

    269_Figure-02

    Figure 2. Olfactory bulb dissection

    Step 4

    Collect the frontal cortex right after the second cut, using a blade and a Dumont No. 5 forceps. Flash freeze the specimen in liquid nitrogen and store at -80°C.

    269_Figure-03

    Figure 3. Frontal cortex dissection

    Step 5

    Collect the striatum (caudate nucleus) right after the third cut with the help of two Dumont No.5 forceps. Flash freeze the specimen in liquid nitrogen and store at -80°C.

    269_Figure-04

    Figure 4. Striatum (caudate nucleus) dissection

    Step 6

    To collect the hippocampus, put the ventral side of the brain up and then remove midbrain to expose the hippocampus. Dissect the hippocampus from the cortex using two Dumont No.5 forceps. Flash freeze the specimen in liquid nitrogen and store at -80°C.

    269_Figure-05

    Figure 5. Hippocampus dissection

    Subscription Required. Please recommend JoVE to your librarian.

    Discussion

    Different brain regions associate with different neurodegenerative diseases. For example, frontal cortex and striatum relate with Parkinson s disease while hippocampus relates with Alzheimer's disease. Precise micro-dissection of the brain allows us to detect the mRNA and protein changes in diseased region more accurately.

    Subscription Required. Please recommend JoVE to your librarian.

    Disclosures

    The animals were handled according to the protocol for the use of animal in research approved by Committee on the Use of Live Animals in Teaching and Research (CULATR) in the University of Hong Kong and according to the National Institutes of Health guidelines for animal use.

    Materials

    Name Company Catalog Number Comments
    Microscope Olympus Corporation
    Dissection tools Metal plate, forceps, blade, etc. Individually wrapped with aluminum foil and heated at 180oC for eight hours to eliminate RNase. Made ice cold before use.

    References

    1. Palkovits, M. Maps and guide to micro-dissection of the rat brain. Elsevier New York (c.1988).
    2. Swanson, L. W. Brain maps: structure of the rat brain. 3rd revised edition. Elsevier Academic Press (2004).

    Comments

    9 Comments

    thank u very much. good protocol
    Reply

    Posted by: AnonymousJanuary 8, 2009, 4:37 AM

    very nice and great
    Reply

    Posted by: AnonymousFebruary 11, 2009, 1:27 PM

    Approximately how much time/rat from sacrifice to remove the frontal cortex and the hippocampus?  Thank you.
    Reply

    Posted by: AnonymousFebruary 16, 2009, 10:59 AM

    Hi, can be done within 3 minutes. When you practice more, ² minutes will be enough.
    Reply

    Posted by: kin c.March 28, 2012, 9:28 PM

    Great, Thanks for your sharing̀²; maybe it's better to have more brain regions to dissect  
    Reply

    Posted by: AnonymousApril 1, 2009, 8:47 AM

    How dŒs this protocol change if we were to attempting to dissect the striatum of embryonic rats or mice for culture?
    Reply

    Posted by: AnonymousJuly 14, 2010, 1:59 PM

    Yeah nicely done. Although I am using mice this seems to work just fine for me as well.
    Reply

    Posted by: AnonymousMarch 28, 2012, 6:23 AM

    Thanks. I think this is just a demostration that we can dissect the brain into regions. When we deal with different parts, normally we go back to the atlas first (you can use the atlas listed in the reference). Then we try to identify the region in the brain samples.
    Reply

    Posted by: kin c.March 28, 2012, 9:26 PM

    Thanks. If we want to isolate some specific nuclei from brain to study them under microscope how we must do that?
    Reply

    Posted by: Jamal E.May 8, 2012, 2:10 PM

    Post a Question / Comment / Request

    You must be signed in to post a comment. Please or create an account.

    Metrics

    Waiting
    simple hit counter