Single-cell RT-PCR as a tool to study gene expression in central and peripheral autonomic neurones

Jacqueline K. Phillips, Janusz Lipski*

*Corresponding author for this work

Research output: Contribution to journalReview articlepeer-review

40 Citations (Scopus)

Abstract

In studies of the central and peripheral autonomic nervous system, it has become increasingly important to be able to investigate mRNA expression patterns within specific neuronal populations. Traditionally, the identification of mRNA species in discrete populations of cells has relied upon in situ hybridization. An alternative, relatively simple procedure is 'multiplex' reverse transcription-polymerase chain reaction (RT-PCR), conducted on single neurons after their in vitro isolation. Multiplex single-cell RT-PCR can be used to examine the expression of multiple genes within individual cells, and can be combined with electrophysiological, pharmacological and anatomical (retrograde labelling) studies. This review focuses on a number of key aspects of this approach, methodology, and both the advantages and the limitations of the technique. We also provide specific examples of work performed in our laboratory, examining the expression of alpha2-adrenergic receptors in catecholaminergic cells of the rat brainstem and adrenal medulla. The application of single-cell RT-PCR to future studies of the autonomic nervous system will hopefully provide information on how physiological and pathological conditions affect gene expression in autonomic neurones. Copyright (C) 2000 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)1-12
Number of pages12
JournalAutonomic Neuroscience: Basic and Clinical
Volume86
Issue number1-2
DOIs
Publication statusPublished - 28 Dec 2000
Externally publishedYes

Keywords

  • α-adrenergic receptors
  • Adrenal chromaffin cells
  • C1 neurones
  • Catecholamine biosynthetic enzymes
  • Multiplex RT-PCR
  • Rostral ventrolateral medulla

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