Neuronal type-specific gene expression profiling and laser-capture microdissection

Methods Mol Biol. 2011:755:327-43. doi: 10.1007/978-1-61779-163-5_28.

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).

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Brain / metabolism
  • Brain / pathology
  • Dopamine / metabolism
  • Female
  • Gene Expression Profiling / methods*
  • Humans
  • Immunoenzyme Techniques / methods
  • Lasers*
  • Male
  • Microdissection / methods*
  • Microtomy / methods
  • Middle Aged
  • Neurons / metabolism
  • Neurons / pathology*
  • Nissl Bodies / metabolism
  • Oligonucleotide Array Sequence Analysis / methods
  • Pyramidal Cells / metabolism
  • Pyramidal Cells / pathology
  • RNA / genetics
  • RNA / isolation & purification
  • Staining and Labeling / methods
  • Young Adult
  • gamma-Aminobutyric Acid / metabolism

Substances

  • gamma-Aminobutyric Acid
  • RNA
  • Dopamine