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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
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 755)

Abstract

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 

Notes

Acknowledgments

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

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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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