Neuronal Type-Specific Gene Expression Profiling and Laser-Capture Microdissection

  • Charmaine Y. Pietersen
  • Maribel P. Lim
  • Laurel Macey
  • Tsung-Ung W. Woo
  • Kai C. SonntagEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 755)


The human brain is an exceptionally heterogeneous structure. In order to gain insight into the neurobiological basis of neural circuit disturbances in various neurologic or psychiatric diseases, it is often important to define the molecular cascades that are associated with these disturbances in a neuronal type-specific manner. This can be achieved by the use of laser microdissection, in combination with molecular techniques such as gene expression profiling. To identify neurons in human postmortem brain tissue, one can use the inherent properties of the neuron, such as pigmentation and morphology or its structural composition through immunohistochemistry (IHC). Here, we describe the isolation of homogeneous neuronal cells and high-quality RNA from human postmortem brain material using a combination of rapid IHC, Nissl staining, or simple morphology with Laser-Capture Microdissection (LCM) or Laser Microdissection (LMD).

Key words

Laser-capture microdissection Laser microdissection Postmortem brain Immunohisto­chemistry Nissl staining Dopamine Parvalbumin GABAergic Pyramidal Neurons Expression profiling 



This work was supported by NIH grants MH080272 and MH076060 (Woo) and R21NS067335 (Sonntag).


  1. 1.
    Sonntag, K. C. (2010) miRNAs and deregulated gene expression networks in neurodegeneration. Brain Research 1338, 48–57.Google Scholar
  2. 2.
    McClain, K. L., Cai, Y. H., Hicks, J., Peterson, L. E., Yan, X. T., Che, S., and Ginsberg, S. D. (2005) Expression profiling using human tissues in combination with RNA amplification and microarray analysis: assessment of Langerhans cell histiocytosis Amino Acids 28, 279–90.Google Scholar
  3. 3.
    Pietersen, C. Y., Lim, M. P., and Woo, T. U. (2009) Obtaining high quality RNA from single cell populations in human postmortem brain tissue J Vis Exp. 6;pii: 1444.Google Scholar
  4. 4.
    Simunovic, F., Yi, M., Wang, Y., Macey, L., Brown, L. T., Krichevsky, A. M., Andersen, S. L., Stephens, R. M., Benes, F. M., and Sonntag, K. C. (2009) Gene expression profiling of substantia nigra dopamine neurons: further insights into Parkinson’s disease pathology. Brain 132, 1795–809.CrossRefGoogle Scholar
  5. 5.
    Simunovic, F., Yi, M., Wang, Y., Stephens, R., and Sonntag, K. C. (2010) Evidence for gender-specific transcriptional profiles of nigral dopamine neurons in Parkinson disease. PLoS One 5, e8856.CrossRefGoogle Scholar
  6. 6.
    Charboneau, L., Paweletz, C. P., and Liotta, L. A. (2001) Laser capture microdissection Curr Protoc Cell Biol Chapter 2, Unit 2 5.Google Scholar
  7. 7.
    Edwards, R. A. (2007) Laser capture microdissection of mammalian tissue. J Vis Exp, 309.Google Scholar
  8. 8.
    Benes, F. M., Lim, B., Matzilevich, D., Walsh, J. P., Subburaju, S., and Minns, M. (2007) Regulation of the GABA cell phenotype in ­hippocampus of schizophrenics and bipolars. Proc Natl Acad Sci U S A 104, 10164–9.CrossRefGoogle Scholar
  9. 9.
    Bartel, D. P. (2009) MicroRNAs: target recognition and regulatory functions Cell 136, 215–33.Google Scholar
  10. 10.
    Barbato, C., Giorgi, C., Catalanotto, C., and Cogoni, C. (2008) Thinking about RNA? MicroRNAs in the brain. Mamm Genome 19, 541–51.CrossRefGoogle Scholar
  11. 11.
    Schratt, G. Fine-tuning neural gene expression with microRNAs (2009) Curr Opin Neurobiol 19, 213–9.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Charmaine Y. Pietersen
    • 1
  • Maribel P. Lim
    • 1
  • Laurel Macey
    • 2
  • Tsung-Ung W. Woo
    • 1
  • Kai C. Sonntag
    • 2
    Email author
  1. 1.Laboratory of Cellular Neuropathology, Department of PsychiatryMcLean Hospital, Harvard Medical SchoolBelmontUSA
  2. 2.Department of PsychiatryMcLean Hospital, Harvard Medical SchoolBelmontUSA

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