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

In studies of the central and peripheral autonomic nervous system, it has b ecome increasingly important to be able to investigate mRNA expression patt erns within specific neuronal populations. Traditionally, the identificatio n of mRNA species in discrete populations of cells has relied upon in situ hybridization. An alternative, relatively simple procedure is 'multiplex' r everse transcription-polymerase chain reaction (RT-PCR), conducted on singl e 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 anatom ical (retrograde labelling) studies. This review focuses on a number of key aspects of this approach, methodology, and both the advantages and the lim itations of the technique. We also provide specific examples of work perfor med in our laboratory, examining the expression of alpha2-adrenergic recept ors in catecholaminergic cells of the rat brainstem and adrenal medulla. Th e application of single-cell RT-PCR to future studies of the autonomic nerv ous system will hopefully provide information on how physiological and path ological conditions affect gene expression in autonomic neurones. (C) 2000 Elsevier Science B.V. All rights reserved.