The isolation of pure populations of neurons by laser capture microdissection

methods and application in neuroscience

Renée Morris*, Prachi Mehta

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

In mammals, the central nervous system (CNS) is constituted of various cellular elements, posing a challenge to isolating specific cell types to investigate their expression profile. As a result, tissue homogenization is not amenable to analyses of motor neurons profiling as these represent less than 10% of the total spinal cord cell population. One way to tackle the problem of tissue heterogeneity and obtain meaningful genomic, proteomic, and transcriptomic profiling is to use laser capture microdissection technology (LCM). In this chapter, we describe protocols for the capture of isolated populations of motor neurons from spinal cord tissue sections and for downstream transcriptomic analysis of motor neurons with RT-PCR. We have also included a protocol for the immunological confirmation that the captured neurons are indeed motor neurons. Although focused on spinal cord motor neurons, these protocols can be easily optimized for the isolation of any CNS neurons.

Original languageEnglish
Title of host publicationLaser capture microdissection
Subtitle of host publicationmethods and protocols
EditorsGraeme I. Murray
Place of PublicationNew York, NY
PublisherHumana Press Inc.
Chapter12
Pages223-233
Number of pages11
Volume1723
ISBN (Electronic) 9781493975587
ISBN (Print)9781493975570
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1723
ISSN (Print)1064-3745

Keywords

  • Brain-derived neurotrophin factor (BDNF)
  • Gene expression
  • High-quality RNA
  • Laser capture microdissection (LCM)
  • Motor neurons
  • Spinal cord
  • Tropomyosin receptor kinase B (TrkB)

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  • Cite this

    Morris, R., & Mehta, P. (2018). The isolation of pure populations of neurons by laser capture microdissection: methods and application in neuroscience. In G. I. Murray (Ed.), Laser capture microdissection: methods and protocols (Vol. 1723, pp. 223-233). (Methods in Molecular Biology; Vol. 1723). New York, NY: Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7558-7_12