Method Article

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

DOI:

10.3791/269

⸱

August 30th, 2007

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

In This Article

Summary

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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.

Abstract

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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

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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.

Rat brain dissection diagram, showing corpus callosum and fornix with labeled cuts for study.

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.

Olfactory bulb dissection, labeled diagram of brain showing olfactory bulb before and after removal.

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.

Frontal cortex dissection process, labeled diagram, for neuroscience educational use.

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.

Dissection of rat brain striatum, identifying tissues; scientific research in neuroscience.

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.

Brain dissection; hippocampus extraction; experimental setup; neuroscience research process.

Figure 5. Hippocampus dissection

Discussion

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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.

Disclosures

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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

List of materials used in this article
NameCompanyCatalog NumberComments
MicroscopeOlympus Corporation
Dissection toolsMetal 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

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  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).

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Tags

RNA ExtractionOlfactory BulbFrontal CortexStriatumHippocampusLiquid Nitrogen

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